How illiteracy influences people’s life Essay Example for Free

How analphabetism influences the great unwasheds life EssayIn the movie, The Reader, Kate Winslet played the role of Hanna Schmitz, an illiterate train conductor who was move to jail without stock-still understanding what she has done wrong. When Hanna said the line, I would rather listen to you , I was moved because these few words allowed me to empathize with people who atomic number 18 illiterate. It gave me a glimpse of the feelings of helplessness and frustration they must encounter in their everyday life. The movie also showed me how illiteracy stern affect a persons life. Because of the point that they do not have access to the information the rest of the world has, they end up being enslaved by the opinions of the literate. It is impossible for them to construct opinions as they do lack the details needed to create one. At the corresponding time, they ar forced to be dependent on people who can provide them with this information. They are also under the mercy of pe ople who have knowledge in the written law. The movie also showed me how humbling it must be not to be able to read or write. At the start of the movie, she had an affair with a man half her age.And, whenever the man would read for her, her seniority faded a counseling because of the way she would act like a child waiting for her father to read her a bed time story. Hannas story shows how principal(prenominal) being literate is. It plays an important role in a persons life as it can change his or her destiny. In this essay, I would like to talk about the changes and effects of literacy to a persons mind and life. I would like to show how literacy can give intellect and great force-out to a person by going through the works of Staple, Douglass and Kozel which they created in order to emphasize the importance of literacy in the democratic world.Brent Staple, an author and editorial writer for the New York Times, recounted the way his grandfather changed his life by acquire how to read. In one of his articles, he wrote Name of Author 2- Nevertheless, the ability to read and write gave them a vantage point on their circumstances and protected them from swindlers who on a regular basis stripped illiterate people of land and other assets. (p. 1) Staples grandfather was able to gain his independence by learning how to read and write. He also versed how to distinguish what is good from what is bad, what can benefit him and what can ruin his life.He also found himself with the power to think and give his opinion as he now has a better grasp of the issues around him. Armed with the knowledge he was able to acquire from reading, he found the power to make his cause decisions and fight for his rights. He was able to free himself from the prison of other peoples thoughts. Another writer, Frederick Douglass, who is famous for his line I would unite with anybody to do right and with nobody to do wrong, wrote an article that is parallel to Staples article in the way t hat he talked about the way literacy influenced his life and his thinking.Douglass story, however, is quite different from that of Staples grandfather. In his story, he recounted that he well-read how to read in secret with the fear of being beaten. Being born a slave, Douglass was passed on from one owner to another. The wife of one his owners taught him the alphabet even though she knew that thither was a law against teaching slaves how to read. Her husband obviously disapproved about this when he found out. He look atd that a slave who gains literacy will be unsatisfied with his condition and yearn for freedom.Even with the threat of being caught and beaten up, Douglass learned how to read with the help of the white children in the neighborhood. He also did his outperform to observe the way the order white men would write. His perspective changed after he has attained literacy. Douglas established that bleak men are not brutes or savages who should be limited to working as a slave, Name of Author 3 they have the right to be educated and respected. As his master predicted, he yearned for the freedom that the white people have. As he continues to learn how to read and write, Douglass attitude to life is gradually transformed by literacy.Douglass mentions in the article I read them over and over again with unabated interest. They give tongue to interesting thought of my own soul. The moral which I gained from the dialogue was the power of truth over the conscience of even a slaveholder. (p. 45) By learning how to read and write, Douglass learned not only the meaning of words and sentences, but the power of knowledge. He slowly became aware of his rights as a human being and how unfairly the black people have been treated. He also realized that illiteracy is the reason why the white people were able to enslave his race for so long.Not only did illiteracy keep them unaware of their rights, it also do them unable to think for themselves, to distinguish f rom right and wrong, what is harmful and what is not. It also stopped them from acquiring the knowledge that they need to pursue their freedom. Hence, they were influenced by the white people to believe that their only purpose in life was to work for them. With his newfound knowledge, he was able to change his life and gain his freedom. At the same time, having appreciated the foster of literacy, he passed on the knowledge to other slaves, giving them the opportunity to change their own lives.In Kozols article, he focuses on how illiteracy can peril a persons way of life and thinking. He believes that one will never really have ability to protect himself if he is illiterate because he is unceasingly unaware of whats going on. He cant read the strange signs on the street that warns him of danger or tells him that he is breaking the law. Neither can he read legal documents, making him vulnerable to being swindled. He is also unaware of his rights. Kozol Name of Author 4 says, Th ey do not know what rights they have, what deadlines and requirements they face, what survival they might choose to exercise.They are half-citizens. Their right exist I print but not in fact. (52-53) They cannot protect their own rights since they dont know what their rights are. Neither can they apply their rights or make a choice because they do not know what their options are. The same points were tackled by Douglas in his article when he said that black people were treated like slaves because they were not literate and were not given the chance to know that they do not have to limit themselves to such a status. They just thought that would go through this nightmare for the rest of their lives because no one could save them.What they didnt know was they couldve saved themselves had they known that they had rights to fight for. If they had been literate, they would have realized that they are not slaves and they have right to be educated and respected because they are humans. Il literacy, however, has exiled them to such an nightmarish existence where their destiny is dictated by their masters. promptly literacy still plays the most important role in society. If you are not able to read and write, it would be really hard for you to live. You wouldnt know how to deal with a lot of things.You wouldnt know what is right or wrong, what would benefit you and what would harm you. Because of the numerous things that you do not know, your lack of knowledge can bring you harm. At the same time, your lack of knowledge, stops you from having your own beliefs or opinions. And, since you have know access to the options you can choose from, you end up believing that there are none and the only thing you can do is follow what the other people are doing or to have the same opinion as they do. Without literacy, all the people can do is follow and obey, allowing other people to take charge of their own lives.If a country is filled with illiterate people then democracy is u seless since the mindless majority can be controlled by the Name of Author 5- government and the votes of those who are literate and have their own minds can be surpassed by those who arent. If that is the case, then the government can become a dictator in the guise of democracy. Hence, it is safe to say that without the presence of literacy, there is no knowledge. Without knowledge, there are no choices. And, without knowledge, there is no freedom which is the very essence of democracy.

Pre-1914 literature Arthur Conan Doyle Essay Example for Free

Pre-1914 litearned run averageture Arthur Conan Doyle EssayThe previous pair of quotes significantly carries the blue(a) assumption of evil and mysterious foreigners. square-toeds associated drugs with the far East As I entered, a swallow of Malay attendant, and the rascally Lascar who runs it has sworn to have vengeance upon me. Both these quotes suggest similar nature in the victorian attitude to others of different origins. The drugs and alcohol is unfairly associated with Malaysia being a Far Eastern country, this unfairly creates unnecessary judgements about the atmosphere of the country. Lascar was a puritanical term used to label an East Indian (Pakistani) sailor army servant. The Lascar in The Man with the Twisted Lip was described as rascally Lascar, although this indicates that the den runner is Asian but also the description creates an image of a mischievous person. We have seen how Arthur Conan Doyle has used stereotypical thoughts in his sound to satisfy the a ristocracy. The Victorians feared that the gloss and customs of non British population, they felt threatened by foreigners Sherlock Holmes was a method of reassurance in which the Victorian culture and values were still respected.During the Victorian era there were three sets of class between people. The highest class were called the Aristocracy, the middle class were called the Merchants, and the lowest class were known as the scant(p). The aristocracy lived a life of luxury and had huge amounts of materialistic wealth and possessions. The aristocracy were all educated and had a high status amongst all people. The merchants could have possibly been literate, but they were not as highly renowned compared to the aristocracy, but they were better off compared to the poor. The merchant class would do a certain job for a living.Lastly, the poor were very nescient and lived in awful conditions. They were detested by the aristocracy because of their lack of hygiene and also because of their class. The aristocracy at the time of the Victorian era were seen as the around trenchant, in The Speckled Band we are make believely shown that Holmes is of the f number class Very soon to knock you up Watson he uses upper class speech. Holmes in The Speckled Band also shows his intelligence No, but I observe the second half of a return ticket this quote shows the wise and intelligent remarks Holmes uses.The upper class were also shown to be very respective and polite, in A instruction in Scarlet, Holmes shows his upper class etiquettes His eyes fairly glittered as he spoke, and he put his hand over his heart and bowed, as it to some applauding crowd conjured up by his imagination. The upper class would foster and incessantly relate to each other, in The Noble Batchelor I presume they were hardly from the same class of society, No I am descending, My last client of the come apart was a king.Holmes tells a client that he has goneureed for a King. In another Sherlo ck Holmes novel The Adventure of Black Peter he frequently refused his help to the powerful and wealthy where the problem made no appeal to his sympathies. These quotes show how Holmes who is of the upper class helps the upper class only if the problem is actualy a logical one for him to have nay interest in. In the customs of Victorian culture Victorian gentlemen were intelligent and rational thinkers and also family leaders and role models.If a man of the upper class fails to acquire these fundamental aspects of a well character mankind they would be known as a fallen gentleman. In The Speckled Band Dr Roylott is a arrant(a) example of a fallen gentleman he beat his native butler to death and narrowly get a instruction a capital sentence, he shut himself up in his house and seldom came out save to indulge in ferocious quarrels with whoever might itch his path. Violence of temper approaching to mania. Dr Roylott is a fallen gentleman, some aspects which make him one is that he is a murderer and has a violent nature.Another perfect example of a fallen gentleman can be found in The Man with the Twisted Lip, the character Mr St. Clair displays clear characteristics of a fallen gentleman to my relief, that instead of being identified as Mr Neville St. Clair, I was arrested as his murderer. Mr St. Clair has been caught acting as a beggar to earn money when he seemed to be a of the higher class standards. Doing such things was an obvious big offence to the Victorian customs.The aristocracy thought of themselves as supreme and pure, they never did any physical work that could discolour their skin a pale face (From The Man with the Twisted Lip). The aristocracy tend to have very pale skin, indicating that they do not work, because work can causes skin discolouring. Hygiene was a very important aspect of everyday aristocracy life, in The Speckled Band, it is very unusual that Holmes packs a toothbrush when he is off to investigate an incident That and a toothbru sh are, I think, all that we need.As well as hygiene, appearance was obviously a very important item addressed in everyday upper class life. From The Man with the Twisted Lip we are shown such as he had started to townspeople in, he has on neither collar nor necktie. This quote reveals to us how important this was to the upper class. This is demonstrating that a man of high class would never be seen without a tie around his neck. The merchant class were not as intelligent and as respected as the aristocracy.The middleclass were not better than the upper class. As reassert by the following quote from The Scandal in Bohemia, it displays a very basic outline in what Holmes does he was still, as ever, deeply attracted by the study of crime, and occupied his immense faculties and extraordinary powers of observation in following out those clues and clearing up these mysteries, which had been abandoned as hopeless by the official police. non only does this quote demonstrate the powers and qualities of Holmes, but in meaning it also shows us that the police who are the working class are being outsmarted by Holmes who is of the upper class, which suggests that the upper class is better than the merchant or working class. The aristocracy also control the middleclass if you leave it to a court of law to clear the matter up on the other hand, if you convince the police authorities that there is no possible case against you (From The Man with the Twisted Lip).This quote shows us the Victorian thinking that a person belonging to the aristocracy will always have power and manipulation over the lower classes. The poor were dislike by the upper classes. The higher class hated them for many reasons, one being there appearance, and extract from The Man with the Twisted Lip He was, as the inspector said extremely dirty, but the grime which covered his face could not c one timeal its repulsive ugliness. This reveals that the Victorian upper class thought the poor as being very u gly, disgusting and dirty, this was the stereotype that all poor people were ugly and dirty.The lower class were seen as the worst of all people the gypsies were hated just because they lived in a different way. In The Speckled Band, the Ms Stoner states sometimes it may have referred to some band of people, by chance to these very gypsies in the plantation might have suggested the strange adjective she used. This demonstrated the class differences, Ms Stoner feels superior to the gypsies, also revealing that the Victorian aristocracy though themselves as being superior to all other classes.Victorian assumption to crime mysteries is that if there was a person out of the upper class or the poor they were the criminals. From The Man with the Twisted Lip, this attitude is clearly shown wash his hands, and his face is as black as a tinkers. Well once his case is settled he will have a regular prison bath. This quotes significance is that the poor people were always the assumed of commi tting crimes, because that is the only way they can survive. When someone decides to change their class it is seen as the worst possible thing that could happen in the view of Victorians.The poor cannot be changed to the aristocracy because of their past and their low status. The same way a man of the aristocracy cannot become lower than his usual class, if this does happen then this is seen as the most shameful situation. vizor examples are Dr Roylott from The Speckled Band and Mr St. Clair from The Man with the Twisted Lip. From analysing mainly the two novels The Speckled Band and The Man with the Twisted Lip that were both written by Arthur Conan Doyle, the Victorian attitudes towards gender, ethnicity and class are revealed.In terms of gender these points have been discussed and proven Men were fare more intelligent and rational than women. Women were over unrestrained they also had authority over domestic matters. However, men were in control over all aspects of Victorian li fe. Points concerning ethnicity were Victorians felt threatened and also hated foreigners (Xenophobia). Victorian saw foreigners and animals as dangerous and mischievous. Victorian towards class was that the Aristocracy were very wealthy, the working class worked for their wealth and the lower class was just poor.Someone transferring classes was not acceptable in Victorian society, the Victorians held a strong belief that someone should stick to their status and class. From the previous Victorian assumption we can conclude that the Victorian morals, values and thinking were much contrasting than what the present views on society are, and things have changed in a positive way. Show preview only The to a higher place preview is unformatted text This student written piece of work is one of many that can be found in our GCSE Arthur Conan Doyle section.

Enterprise Resource Planning Systems For Smes

Enterprise Resource Planning formations For SmesSeveral researches score revealed that Enterprise Resource Planning (ERP) systems take away greater realises to transcriptions, sm t turn out ensemble or large. These reports asserted that successful weaponation of the right ERP tooth root will empower the monetary success of the formation while its spliture throw out spell doom and gloom. SMEs in contrast to their larger counterpart choose peculiar challenges that further make the choice of ERP dissolving agent difficult. These challenges shit defined the expectations and requirements of SMEs from ERP solutions. several(a) ERP softw are package are getable in the foodstuff for their use, with the software leader being patsy, scarcely it is essential for an SME to select the appropriate software solution that suitably address their requirements putting in consideration their peculiar predicament. This is easily achieved through an adequate software selection and ev aluation process. This research looks into past kit and boodle on ERP lend oneselfations for SMEs to come up with ERP requirements for SMEs available software (and hardware) solutions available to them and a detailed software evaluation and selection process in line with industry best practice which they elicit use. It also addresses current ERP market and market trends.MethodologyThis paper is largely sensuald on re receive of previous schoolman documents in form of journals, conference documents and books. These were reviewed extensively in line with the topic of discussion. Another helpful source of breeding used in the report is the internet. with the internet, we were able to get more current information and data to support previous academic works.While we prevail d whizz extensive research on this topic, it should be noted that the academic documents used in this research represent just a subset of the numerous documents that have been written concerning this topic. T hey are by no means a perfect representation of all the possible views.Table of limit1Abstract 1Methodology 1Table of Contents 2Introduction 3Enterprise Resource Planning (ERP) Systems 4 micro and Medium Size Enterprises (SMEs) 4ERP Market and Market Trend 6ERP Requirements for SMEs 9Analysis of ERP resolution (Software and hardware) 11 s buttocks 11 prophesier ERP Software 14International Financial service (IFS) 15SAGE Accpac 15Microsoft Dynamics 16Open starting time Software (OSS) 16ERP Evaluation and Selection process 17 stop 1 Plan Requirement 18 put 2 Request for Proposals (RFP) 19 map 3 ascendant Evaluation 19Stage 4 Contract Negotiation 19Stage 5 Selection and scoffment 20IntroductionEnterprise resource planning (ERP) systems, gibe to Joseph et al 2003, is now being promoted as a lovable and critical link for enhancing integration between all functional areas at heart an try and between the enterprise and its upstream downstream trading partners.Despite all ben efits potentially offered by ERP systems (Banker, 1998 Hicks, 1995 Minahan, 1998) however, the issue of organisational qualifyings needed by ERP implementation as wholesome as predicting return on investment assumes particular importance within small and medium-sized enterprises (SMEs) because of their peculiar challenges.In SMEs, the implementation and the evaluation of the potential benefits is still a difficult task. (Ravarini, 2000).(Chan, 1999), asserted that many SMEs either lack enough resources or are not enthusiastic about committing a huge component of their resources to ERP implementation due to long implementation period and last fees associated with ERP, (Chau, 1995). These amongst other factors have resulted in slow ERP adoption by SME (Tagliavini et al, 2002).Nowadays, some SMEs are not only seeking ways of integrating their various island-of-information within the organisation, but have also moved to ex thin the whole scope beyond their organisation to implicat e their suppliers, trading partners and customers (Charlesworth et al., 2002). Therefore, implementation of a right and cost effective ERP solution is essential. This however must be preceded by an accurate ERP software evaluation and selection process.This paper explores ERP for SMEs and is formatted as follows Introduction, Methodology, Background on ERP, ERP Market and Trends, ERP and SMEs, ERP requirement for SMEs, distinguishable Software (and Hardware) Solutions Analysis a Detailed Software Evaluation and Selection Process, Discussion, Conclusion and Future Research work.Enterprise Resource Planning (ERP) SystemsERP systems butt end be defined as the intimately predominant computing systems for blood in a lot of organisations, both in the public and private sectors (Gable, 1998)The ERP software can be customised to support critical existing processes which organisations follow (Stefanou, CJ., 2001)Included in an ERP system are Storage, orders, assets, vendor contacts, pur chasing, financials and human resources (Dahlen C. and Elfsson J., 1999)(OLeary, 2000) stated that An ERP system amalgamates the majority of business processes and allows access to the data in real time. However, (Gardiner et al., 2002) also stated that an ERP system improves the performance level of a supply chain by helping to shrivel up cycle times.Other benefits which enterprises enjoy by implementing an ERP system include better customer satisfaction, reduced quality cost, increased flexibility, change resource utility, improved purpose-making capability, improved information accuracy and improved vendor performance (Siriginidi, 2000).The flip-side to this is that, most ERP software tend to be complicated, large and costly (Mabert et al., 2001). The implementation of an ERP system is time overpowering and puts pressure on an organisations information applied science (IT) department or outside professionals (Khaled Al-Fawaz et al., 2008)Putting ERP in place requires new p rocedures, employee training, and both fill outrial and technical support. (Shang and Seddon, 2002)Small and Medium Size Enterprises (SMEs)SMEs, in most countries are businesses that employ fewer than 250 people. For some other countries, SMEs are those which employ fewer than 500 people. Countries like these include USA, Italy and Australia. 99.9% of the businesses in Albania come under SMEs. (Nito E., 2005)ERP customers are usually split into terce market air divisions by annual income. These include large, midmarket and small. The borderline of these divisions tend to be different, depending on which ERP vendor is victimisation them.For small enterprises, Sage, consume, Microsoft, Infor and to a slight extent Lawson, compete with a number of smaller vendors for ERP systems.Figure 1 Source bla bla blaAs shown in the diagram above, only SAP and Sage have set ERP returns designed for the small enterprise market. The other vendors compete in this division with growths that the y also use for the midmarket.The top six vendors compete in the mid-market. A lot of these products are industry specific e.g. pharmaceutical distribution, drop point services, process manufacturing, etc. Successful competition has been attained by the leading ERP vendors adding sector specific focus to their products.This has been approached by Industryspecific preconfigured templates using a standard ERP product being developed by the ERP vendor (e.g. Oracles Business Accelerators or SAPs Best Practice Templates).ERP Market and Market TrendTrend, as defined by (Kotler, 1997) is A direction or sequence of events that have some momentum and durability.Over the last decade, the ERP market has grown very quickly. The three main reasons for this growth are internationalisation, year 2000, and the need for better information integration. (Dahlen C. and Elfsson J., 1999)As at 1998, Merrill Lynch estimated that 40% of companies with revenue enhancements over $1billion had already imple mented base ERP systems in the USA (Cald wholesome Stein, 1998) and had started implementing additional applications, a market that was estimated at $8billion by 2002.An American-based research firm, AMR Research, also predicted the enterprise applications market could hit $78billion by 2004 compared to $27million in 1999 (Stefanou, CJ., 2001). However, according to Gartner group, a high percentage (88%) of the ERP market is concentrated in the western countries.SAP, the German flagship go with, holds a global market share of 32%, while Oracle has a global market share of 14.5%. Both vendors have the lion share of the spheric ERP market. However, this is not the eccentric person in the Chinese market where SAP and Oracle hold a combined market share of 24.4% and the domestic companies holding a combined market share of 51.6%. (Huigang Liang et al., 2004)Total ERP market revenue grew to over $28B in 2006. This was driven by a combination of strong customer demand and leading vend ors expanding their product portfolios through acquisition. (Jacobson S., 2007)As at 2000, the AMR Research, Inc. predicted that the ERP market would grow at a compound annual growth rate of 37% over the following five years.Currently, SAP and Oracle have dominated the market. Their competition, however, lies in the SMEs market With rivals like Sage group, Microsoft, Epicor and Lawson (Jacobson S. et al 2007) infra is an ERP application revenue estimate for 2006 2011This was estimated as at 2007 by the AMR, Research GroupFigure 2 (Jacobson, S. et al 2007 The ERP Market sizing Report, 2006-2011, AMR Research., ERP 2007 Market sizing series).In recent times, the ERP Vendor Outlook has changed a lot. Six vendors are responsible for almost two thirds of global ERP sales. Oracle and SAP control 50% of the World market by income. At the start of the last decade, Infor and Microsoft did not have a market presence.Below is a pie chart showing planetary ERP Revenue by Vendor (2008) Figur e 3 Source BSM, ERP. May 2010ERP Requirements for SMEsAccording to Huin 2004, SMEs have an enormous rest in needs, operating requirements, logistics fulfilment and financial capabilities compared to their large counterparts.First among these requirements is a cost effective ERP solution. Costs associated with ERP solution include not only the software, but also to items such as training, hardware and consulting (Willis et al 2001).Besides, most ERP software available in the market, according to Bounanno G. et al 2005, are just too expensive for SMEs.Chan R. 1999, supported that many small-medium enterprises either do not have sufficient resources or are not willing to commit a huge fraction of their resources due to long implementation times and high fees associated with ERP implementation. This emphasises the fact that cost is a major factor for SMEs when attempting to undergo a capital intensive think like ERP implementation. A research of 50 Italian SMEs carried out by Federici 2009 suggested that small and medium enterprises mostly chose systems provided by small national software houses on the face of it for cost reasons.Contrast to larger establishments, SMEs have limited experiences, human resources and skills that are necessary for some kind of ERP implementations according to Adam and ODoherty 2003 cited in Winkelmann and Klose 2008. It is a well known fact that most SMEs have a scaled down number of employees who handles more than one function.This position is further supported by Huin 2004, which confirm that most SMEs experience a high turnover rate compared to large companies. That work further stated that high turnover rate adversely impacts how SMEs manage their labour resource-a key resource input in any ERP project.This is in line with the research work of Winkelmann A et al 2008 which emphasised that a sanctified IT department addressing the selection and implementation of an ERP system dont actually exist in most SMEs. As such most SMEs have requirements for ERP solutions (software and hardware) that leverage on their lean human resources, especially in the IT department and provide a user friendly platform not riddled with so many complications.Achanga et al 2006 substantiate in his work that most SMEs do not have a good instruction structure on which innovations such as ERP thrives. It is customary to have the Managing Director or the CEO involved in daily administration of the companies in addition to making strategic decisions. They further explained that most SMEs were established by owner managers who may not have the tactful management know-how. As such, strategic progress is usually hampered by lack of good leadership traits which bring about informal working processes. This was supported by (Mintzberg H. et al 2003 p.217).Processes here according to Beretta 2004 is the medium that coverts activities efficiently and effectively in order to generate survey for a specific customer. It is important to note that these processes are what ERP serves to integrate. As such ERP solution for SMEs must be simple and flexible to accommodate these peculiarities of most SMEs. such flexibility involves things like allowing only specific modules or even sub-modules to be implemented.The type and focus of the SMEs should begat the requirement for consideration in the choice of ERP solution. Most SMEs have very specific operations and as such have very specific processes. These processes differ according to the specialisation of the company. Example is the case of a look company in Taiwan that wanted to implement ERP but could not get software fit because of its unique operations (Yang J et al, 2007).Another exercising Made-To-Stock (MTS) manufacturing companies have different ERP requirements from Made-To-Oder (MTO) companies (Deep A. et al 2007). The implication is that MTO and MTS have different production processes and thus will need a system that aligns with their peculiar requirement.We can and so conclude according to Deep A. et al, 2007 that issues relating to the specificity of an organisation will need to be determined before proceeding to make a choice on the ERP solution for implementation.Analysis of ERP Solution (Software and hardware)ERP software belongs to a suit of software called Enterprise system software. According to Shanks G. et al 2003, it isa set of packaged application software modules, with an integrated architecture, that can be used by government activitys as their primary engine for integrating data, processes, and information technology, in real time, across internal and outer tax chainsimpound deep knowledge of business practices that vendors have accumulated from implementations in a wide range of client organizations, that can exert considerable influence on the design of processes within new client organizationsis a generic semi-finished product with tables and parameters that client organizations and their implementation partners must configure, customize and integrate with other computer-based information systems to meet their business needs.Thus, ERP software are sold to organisations on the platform that its a way for them to implement best practices in an organisation (Wagner and Newell 2004), but there is a vast gap between this theory, the actual implementation and use of the software. The idea behind the software use is that a immix of best practice within a particular industry is built into the design so that this can be used by a alike(p) company irrespective of where it is to automate its working process.According to Holland and Light 1999, most organizations now opt for off the shelf software instead of developing one in-house for its function.While there are over one hundred ERP software available, we will be reviewing a list of some well known ERP solutions available to an SME belowSAPA company that started in Germany, the application has captured the integrated market of most large and medium mar ket throughout the world (Martin and Cheung 2000). With high functionality and a great deal of integration, the solution covers requirements such as financial accounting and control, sales and distribution, materials management, production planning and human resource management.Bancroft et al 1998 revealed that SAP design consists of the following layers of softwareThe SAP graphical user interface (GUI) representing the presentation layerThe SAP application layer andThe SAP database layerSAP supports critical business functions and processes and can actually be tailored to meet the business needs of any enterprise (SAP Global, 2010). It delivers the following solutionsSAP ERP Financials helps addressing changing financial reporting standards, improve cash flows and manage risksSAP ERP Human Capital centering helps in automating key processes like End-user service delivery, work force analytics, talent management, workforce process management and workforce deploymentSAP ERP Operatio ns helps with process like procurement and logistics execution, product development and manufacturing and sales and servicesSAP Corporate operate covers real estate management, Enterprise asset management, project and portfolio management, travel management, environment and safety management, quality management and global trade servicesSAP Global claimed that the product helps in increasing oversight of business operations and providing adequate information for strategic business decisions. It also claimed high flexibility and innovation and that modules can be implemented as needed and upgraded as the need arises. The SAP for SMEs has been implemented by over 80000 SMEs.Figure 4 below shows the SAP Solution roadmap showing the multiple level of blueprint of processes supported by SAP.SAPs benefits were echoed by the research work of Mandal and Gunasekaran 2003. This was implemented at PMB Water Corporation. They discovered greater benefit of implementing SAP.However, Al-Mashari an d Zairi 2000 confirmed that SAP is a very complex solution that is too prone to failure. This position is supported by the research work of Martin I. et al, 2000 in an organization in Australia where he discovered that the training was complex, expensive and enormous. It is also not a user friendly application. For a small and medium sized company this enormous complexity and capital investment among other things is not one that can be readily and easily accommodated as previously stated in the requirements section.Figure 4 SAP Solution Roadmap (available at http//www. sap.com/businessmapsHowever, it should also be noted that quite a number of medium establishments have been able to implement SAP successfully and are reaping the benefits (Martin I. et al 2000).Oracle ERP SoftwareThe Oracle ERP software is part of the Oracle e-Business suite. It consists of the following modules (Oracle 2010)Channel Revenue Management enables information driven channel management. Includes modules l ike Accounts receivable deductions settlement, channel rebates and point-of-sale, partner management, price protection and supplier ship and debitFinancial Management covers things like addition lifecycle management, cash and treasury management, credit-to-cash, financial control and reporting, financial analytics, governance, risk and compliance, lease and finance management, procure-to-pay and travel and expense managementHuman Capital Management includes Global Core HCM , Workforce Management, Workforce Service delivery, Integrated talent management, and HR analyticsProject Portfolio Management Involves things like project analytics, project billing, project contracts, project collaboration, project costing, project management, project portfolio analysis, project resource management and time and labour.According to Oracle, Oracle E-Business suite is the most adaptable global business platform and the most customer-focused application outline. This opinion seems to be supported b y Panorama Consulting group 2010, that the software has the highest level of predictability of all ERP software that was studied, and that the average initial cost to implement is 25% less than that of SAP. It also claimed that Oracles total cost of ownership is nearly 50% less than that of SAP. The fact remains though that its usage does not as yet compare to that of SAP. This is true considering the fact that there has not been much academic research into the use of the software as an ERP solution.International Financial Services (IFS)IFS is a single, integrated product supporting the management of 4 core processes (IFSWorld 2010)Service and Asset, Manufacturing, Project, Supply chain.It can be added to other suites of software e.g. Financials, Human Resources, gross revenue and services, Engineering, Project, Manufacturing and Distribution. According to IFSWorld 2010, its easy to use software that provides an attractive, intuitive and efficient user experience. It is also quite f lexible, allowing an organisation to choose the required module and upgrade at a future date if so desired.The benefits of IFS was supported by the work of Lahikainen T. et al 2000 comparatively saying that ERP software like SAP is rather unmanageable and large, and because of this cannot easily be reworked to suit a business process. Rather, the business will have to readjust its process for the software.SAGE AccpacThis product offers middle sized organisation an adjustable enterprise resource for finance, SRM and operations (Sage, 2010). It supports global aspirations for mid-market and upper-mid-market companies with its robust financial management capabilities and flexible open architecture.Modules of the software includeFinancials General Ledger, G/L Consolidations, Reporting, Multi-Currency, Intercompany Transactions, Transaction Analysis and Optional Field CreatorPurchasing Purchase Orders, Accounts Payable, Electronic Funds Transfer (ETF), Direct payables, Document Manageme nt, Fixed Asset Management, couple and Form printingSales and Receivables Order Entry, Accounts Receivable, EFT Direct Receivables, Electronic Data Interchange (EDI), National Accounts Management, Unit Sales Analysis, Return Material Authorization (RMA) guest Management Contact Management, Sales Force Automation, Marketing Automation, Customer Service AutomationInventory/Warehouse Management Inventory Control, Lot Tracking, Serialized Inventory, Warehouse Management, Manufacturing ManagementProject Management Project and Job Costing, Service and Maintenance management,Payroll In-house Payroll, Electronic Funds Transfer (EFT), Direct Payroll, Sage Accpac HRMSSystem Administration System Manager, Alerts and Alerts ManagerSage manages the processes that are common to all businesses, such as finance, HR, or CRM the vertical modules give customers a greater ability to match technology to specific industry needs (Sage, 2010).Microsoft DynamicsAccording to Microsoft 2010, Microsoft Dynami cs offer solution that can help fastrack performance, measure financial effectiveness and enhance decision making. It helps drive businesses by providing a backbone of an elastic system. It is easy to implement and adapt. Apart from on-premise installation, the solution can also be deployed to work with cloud computing, a solution some organisations are already considering.The capabilities delivered by this solution includes Financial management, Supply chain management, business intelligence, performance management, Collaboration, Project Management, Human resource management, IT management and Software-plus-Services.Open Source Software (OSS)There are also a number of open source ERP software that seems to be of an increasing interest at the moment according to the research work of Johansson and Sudzina 2008. The list includes Compiere, OpenBravo, Opentaps, Facturalux, TinyERP.It may be that most SMEs will meet OSS ERP implementations able to cater for their needs since according to Raymond 2005, SMEs are highly flexible and adaptable to change. Implementation may also work out cheaper as costs associated with licensing fees are usually not incurred (Johansson and Sudzina, 2008). It should however be noted that support for implementations for OSS ERP software is quite hard, as there can be several versions of a given software and finding an expert can be daunting.ERP Evaluation and Selection processA successful ERP project requires selecting an ERP solution, implementing the solution, managing changes and examining the practicality of the system (Wei and Wang, 2004). Wrong ERP solution choice would either cause implementation to fail or weaken the system to a greater impact on the enterprise (Hicks,1995 Wilson,1994).Most enterprises often jump into looking at ERP functions and features rather than examining the strategy and business processes. According to Donovan, 2001, it is important for management to know the current strategy, processes and supporting s ystems compared to what they could be with the new systems.For most enterprises, the decision to implement ERP functionalities will require buy a software package from one of the more popular vendors on ERP market. But the selection process is not a straightforward task, hence thoroughgoing(a) understanding of what ERP packages are to offer, differences in each of them and what might be at stake in selecting one package over the other should be well examined or evaluated, (Sammon and Adam, 2000).Evaluating and selecting an ERP system, even though can be a very complex process (Donovan 2001). It should be a fact-based process that will bring an enterprise to the point where comfortable well-informed decisions can be made.Figure 5 below shows an online poll conducted by ESI International survey of 2,000 business professionals in 2005. This clear indicated that most software projects, ERP inclusive, failed due to lack of adequate requirements definition.Fig. 5 Source ESI Internatio nal survey of 2,000 business professionals, (2005).In addition to the above figure 5, Donovan pointed out that typical ERP project implementation can also fail because of a wrong choice of ERP solution. Therefore, Management legerity Inc in 2005 revealed that it is imperative to adopt a thorough evaluation and evaluation process before adopting any ERP solution in SMEs. Their report detailed eight steps necessary for a thorough and reasonable level of successful ERP implementation in SMEs. This is represented in Figure 6. We have further categorised this into the following five stages for our discussionPlanning, RFP, Solution Evaluation, Negotiation, Selection AgreementThese five stages are explained below based on the research carried out by Management Agility Inc in 2005.Stage 1 Plan Requirement trace business along with areas of business that require technical approach.Develop a specific business case with business value for a solution.Ensure that the project sponsor is willi ng to articulate the business case for change, identify vendors and get familiar with the available solutions.Get general view of investment needed, considering software, hardware, other related infrastructure and ongoing support.Evaluate the organisation readiness for the investment and decide whether to continue or not.Define priorities under must-have and nice-to-have accordingly.Stage 2 Request for Proposals (RFP)Shortlist interesting vendors based on the outcome of market survey for solutions and then for demonstration.Collects facts in line with the business need from product demonstrations for the development of unbiased RFP for vendors.Set-up a neutral body to develop RFP, using facts gathered from products demonstration aligned to the business requirements.Distribute out RFP to selected vendors.Generate canonical expectations from ideal proposal in line with the business need for onward selection of the ideal software vendor.Stage 3 Solution Evaluation draw and prioritis e remaining gaps between software capabilities as demonstrated and business requirements.Identify how the gaps will be bridged in terms of configuration, process change or a combination of all these.If the gaps cannot be bridge, then discontinue the evaluation exercise immediately, otherwise consider reengineering of those affected business processes and continue with the evaluation.Stage 4 Contract NegotiationNegotiate with each vendor establish software, hardware and other infrastructure agreement requirements, which include version, components, maintenance and support. Also negotiate interlocking in user groups, license costs, maintenance fees and many others.Establish service provider agreement which also include deliverables, timelines, resources, costs, payment schedules and other legal requirements.Stage 5 Selection and AgreementUpon successful negotiation with the right vendorReview all legal terms on privacy protection, operation counsellor and data manipulation etc.App rove agreements with the selected vendors.Agree on implementation plan.Fig. 6 Detailed flowchart for ERP Software, Hardware Evaluation and Selection Processes Management Agility Inc, 2005ERP Software Hardware (Solution) Evaluation and Selection StepsYesNoStage 4Stage 3Stage 2Stage 1Define RequirementsShop Round for ProductClarify RequirementsEvaluation Vendor InquiryInteract with VendorsNegotiate AgreementAction AgreementDefine business case/need and spell-out required values. Be specific. Ensure the business sponsor is willing to push through business case for change.Look round the market for what product is available. Identify vendors that operates and their general approaches to technologies the take. Discuss with others in the same industry as you are etc.Clarify your requirements and be sure of what you are looking for in line with you business case. Refine requirements if possible and be specific too.Find out what product is looking promising in line with the business need an d from which vendor. Identify which vendor and their products and invite interesting ones for demo etc. Request for proposal (RFP).Invite each shortlisted vendor over for a chat and find out more about the product. List out expectations based heavily on business requirements.At this point evaluate this approach. Can you afford to change your current process? Can you afford the change the new product will bring and many more?Initiate Negotiation for the selected product with the selected vendor. Agree on who does what, when are they to be done. Negotiate deliverables, timelines, co

Construction of Recombinant DNA in E Coli

Construction of Recombinant deoxyribonucleic acid in E ColiIn 1973 Stanley Cohen and Herbert Boyer pioneered the use of recombinant deoxyribonucleic acid technology for re-create and musing of genes in foreign organisms. They cloned desoxyribonucleic acid from the Salmonella typhimurium streptomycin resistance plasmid desoxyribonucleic acid RSF1010 into the Escherichia coli plasmid pSC101 and observed tolerance to streptomycin among the transformants (Cohen et al., 1973). The first reported production of a human recombinant protein excessivelyk send off a few years later when the thusly newly inaugurationed biotech company Genentech announced that they had managed to express the gene encoding human somatostatin in E. coli (Itakura et al., 1977). The value of the resulting bio moive nerve center was similar to that of somatostatin extracted from the brains of 500.000 sheep. In 1982 Genentech followed up this success with the product humulin, a recombinant insulin produced i n E.coli and the first recombinant biotech drug to be accepted for market by the Food and do drugs Administration. Today the production of recombinant proteins has become a huge global industry with an annual market volume exceeding $50 billion (Schmidt, two hundred4). At the start of the recombinant protein expression era the bacteria Escherichia coli and Bacillus spp. dominated as entertains for recombinant expression, but the realization that a protein may require a specific host physiology and biochemistry for optimum production stimulated a search for new hosts, both prokaryotic and eukaryotic. Par every(prenominal)el to this quest, recombinant DNA technology advanced tremendously thereby opening up possibilities for the use of novel organisms. As a conchronological succession, many unalike expression systems for use in many different hosts argon now available, including systems for use in yeasts (Gellissen et al., 2005), filamentous fungi (Nevalainen et al., 2005), insect a nd animal cell cultures (Wurm, 2004 Kost et al., 2005), gram-positive bacteria like Bacillus (Westers et al., 2004) and Streptomyces (Binnie et al., 1997), and gram-negative bacteria like Escherichia coliBacterial expression systems atomic subject 18 the preferred choice for production of many prokaryotic and eukaryotic proteins. The reasons for this lie in the cost- gistiveness of bacteria, their well-characterized genetics, and the availability of many different bacterial expression systems. Among the hosts available for recombinant expression, Escherichia coli is in an exceptional position. This stems from the many decades of intense researchon its genetics as well as the broad scope of biotechnological tools available for genetic engineering of this organism. As a host for recombinant expression, E.coli is especially valued because of its rapid growth rate, capacity for continuous fermentation, low media costs and achievable high expression levels (Yin et al., 2007). One cons equence of this popularity is that about 80% of all proteins used to solve three-dimensional structures submitted to the protein data bank (PDB) in 2003 were prepared in E.coli (Srensen and Mortensen, 2005) and during 2003 and 2006, nine out of 31 approved therapeutic proteins were produced in E.coli (Walsh, 2006), among them important growth actors, insulins and interferons (Schmidt, 2004).Green Fluorescent Protein (GFP) was isolated from the jellyfish Aequorea aequorea in 1962 (Shimomura et al., 1962) where it was found as a companion protein to aequorin, the well-known chemiluminescent protein of the same species. It was detect that living A. aequorea tissue had an emission spectrum peaking at 508nm and looking green but pure aequorin peaked in the blue range, at 470nm (Tsien, 1998). This indeed led Shimomuras multitude to discover GFP and suggest radiation-less energy transfer as the mechanism for exciting the protein. Its structure has been designated to consist of an 11 s tranded -barrel containing the chromophore made up of a single helix as shown in Figure1.Its use as a tool in molecular biology was not realised until 1992 when Prasher reported the re-create and sequence of GFP (Prasher et al., 1992). Since 1994 GFP has been used as a reporter protein (Chalfie et al., 1994) flagging its own presence and therefore also proteins under the same experience, by emitting green glitter (em = 508 nm) upon excitation with near ultraviolet light (around 395 nm) or blue light (around 470 nm) (Ito et al, 1999). Since then many mutations have been developed looking to improve the emission or to focus it to a single wave space (Heim et al., 1995) or to change the color of the emitted light itself.Recombinant DNA molecules unremarkably contain a DNA fragment inserted into a bacterial vector.Polymerase chain reaction (PCR), a specific gene or DNA region of interest is isolated and amplified by DNA polymerase extracted from a heat-tolerant bacteria. PCR finds the DNA region of interest (called the target DNA) by the complementary binding of specific short primers to the ends of that sequence. The long chromosome-size DNA molecules of genomic DNA must be cut into fragments of a much smaller size to begin with they can be inserted into a vector. Most cutting is make with the use of bacterial restriction enzymes. These enzymes cut at specific DNA sequences, called restriction sites, and this property is one of the central features that lease restriction enzymes suitable for DNA manipulation. These enzymes are examples of endonucleases that split a phosphodiester bond (Anthony, 2012). The key property of some restriction enzymes is that they make sticky ends. The restriction enzyme EcoRI (from E.coli) recognizes the following sequence of six nucleotide pairs in the DNA of any organism5-GAATTC-33-CTTAAG-5The enzyme EcoRI makes cuts merely between the G and the A nucleotides on each strand of the palindrome (Figure.2).The recombinant DN A molecules are transferred into bacterial cells, and, generally, that one recombinant molecule is interpreted up by each cell. The recombinant molecule is amplified along with the vector during the division of the bacterial cell. This process results in a clone of identical cells, each containing the recombinant DNA molecule, and so this proficiency of amplification is called DNA cloning. The succeeding(prenominal) stage is to find the rare clone containing the DNA of interest.Bacterial plasmids (vectors) are small circular DNA molecules that replicate their DNA independent of the bacterial chromosome. The plasmids routinely used as vectors carry a gene for drug resistance and a gene to distinguish plasmids with and without DNA inserts. These drug-resistance genes try a convenient way to select for bacterial cells transformed by plasmids those cells still alive after exposure to the drug must carry the plasmid vectors. However, not all the plasmids in these transformed cells en trust contain DNA inserts. For this reason, it is desirable to be able to identify bacterial colonies with plasmids containing DNA inserts. Such a feature is part of the pUC18 (or pUC19) plasmid vector shown in Figure 2 DNA inserts disrupt a gene (lacZ) in the plasmid that encodes an enzyme (-galactosidase) necessary to vex a compound added to the agar (X-gal) so that it produces a blue pigment. Thus, the colonies that contain the plasmids with the DNA insert will be white rather than blue (they cannot cleave X-gal because they do not produce -galactosidase).The following experiment outlines the construction of recombinant protein production in E.coli strain BL21 by using a bacterial plasmid vector pUC18/19 expressing Green Fluorescent Protein (GFP) to act as a recombinant protein product with the benefits of being easy to visualise and measure.Materials and MethodsMaterialsThe experiment was carried out using the following materials and Equipments 2l EcoRI/HindIII cut and cleaned PUC19 vector, 5l EcoRI/HindIII cut and cleaned GFP insert, 2l 10xT4 ligase buffer, 2l T4 ligase(0.5 U ml-1) , and 9l sterile water (H2O) to make up to 20l volume . atomic number 6l of competent BL21 E.coli cells on ice, 42C water bath, Ice bucket with ice, selective media plates (1.5% Luria broth (LB) Agar, 40g mL-1 X-gal, .1 mM IPTG, 50g mL-1 ampicillin), sterile tubes, shaking incubator, Spectrophotometer or similar device to measure optical density of the bacterial cultures, flasks, Microcentrifuge.MethodsIt can be divided into three stagesLigation Reaction stage in this stage 2l EcoRI/HindIII cut and cleaned PUC19 vector, 5l EcoRI/HindIII cut and cleaned GFP insert, 2l 10xT4 ligase buffer, 2l T4 ligase (0.5 U ml-1) , and 9l sterile water (H2O) are mixed and kept at room temperature for at least 30 minutes. version of ligation into cloning host stage this stage conducted by deforesting 100l of competent BL21 E.coli cells on ice (with caution do not allow to firm to room temperat ure), then adding 10l of the ligation reaction from the first stage to BL21 E.coli cells. They are then incubated for up to 30 minutes on ice. Next step, is done by taking out the displacement mixture out of the ice and heated in water bath at 42 C for well-nigh 75 seconds, then followed by return direct into ice for a minimum of 2 mins. Then the cells were plated out on selective media plates (1.5% Luria broth (LB) Agar, 40g mL-1 X-gal, .1 mM IPTG, 50g mL-1 ampicillin). Lastly, the transformation mixture is incubated at 37 C for 12-18 hours afterdriedd.Picking of colonies for the protein expression stage 2x5ml LB +50g ml-1 ampicillin in 30ml sterile tubes were prepared, then 1xBlue individual colony and 1x white individual colony selected and inoculated in separate tubes. Then the tubes were incubated with shaking incubator passim the night at 37C , revivify 220rpm.Subculture and Growth of Recombinant E.coli for Protein expression At the beginning, 2x60ml sterile Luria-Bertani (LB), in 250ml conical flask were warmed , (1 per inoculums ) at 37 C, Then aseptically the ampicillin was added to a last stringency of 50g ml-1 ampicillin. Next 1 ml of media was remote and was put in a cuvette to act as blank (one blank is enough for both ouh), followed by addition of 600l overnight to calture of each individual colony to separate flask (1100 inoculum), the flasks were put back to the shaking incubator and incubated at 37C, speed 200rpm , after that blank spectrophotometer was placed against media at 600nm , after 45 minutes the samples were removed aseptically from flasks, then from every flask 1x 1mL was removed and added to a new clean cuvette (take to next step 8) and 1x1ml was added to clean Eppendrof (take to step 9) . The OD600nm of culture in cuvette was Measured and the result of growth curve was recorded (once the culture has reached an OD 600nm of 0.5, IPTG was added to final concentration 1Mm stock solution. Then samples were spun down in the Eepp endrof tube at max speed in Microcentrifuge for 5 minutes , ensure centrifuge is balanced in the lead spinning , the supernatant was removed and pellet ,then the pellet was suspended in 200l Cell lysis buffer (10mMl Tris PH8.0, 300Mm Nacl , 10mg ml-1 Lysozyme). Resuspended cells were frozen at -20 c to the next day. Lastly, sampling was continued until OD600nm is no longer rising for two successive samples or until 1630 pm.Results and discussionAlthough it is supposed to harvest between 30-300 colonies per plate (210- 2100 colonies for all groups), just three blue colonies were observed in plates between all groups, which mean that protein of interest (GFP protein ) was not expressed (inefficient) in BL21 E.coli cells due to some factors influenced the expression level or to some technical problems during the experiment which will be discussed.The most popular strain, BL21 and its derivatives, which are unspoilt producing protein, are descended from E.coli B and thus is deficient in the Lon protease. Additionally, the BL21 background lacks the OmpT outer membrane protease. For expression work, BL21 cells should be taken from stock cultures that performed from fresh transforms. This step is crucial to insure that the clone does not change and that each expression run gives optimal performance.Transformation frequency is bear upon by the purity of the DNA, how the cells are handled, and how the transformation was actually performed. In the impurities in the DNA usually spin columns can be used to purify DNA from PCR reactions, ligations, endonuclease digestions, or other treatments. In addition, the most common mistake when transforming E.coli is to put a lot of ligation mixture in the transformation.Other factors that effect transformation with BL21 are the handling and the storage of the competent cells. Competent cells need to be reserved at -70C to keep them at the peak .It is worthy of noting that 5-10-fold of efficiency usually lost if tube put back in the box and place in the freezer. Moreover, Cells must be thawed on ice, and the transformation should be started immediately after the cells are thawed. Incubating on ice is necessary for chemically competent cells. If you heat shock right away, the efficiencies will be down 10-fold. If incubate for only 15 minutes, it will be down 3-fold. In addition, time of heat shock (75 second) could be not enough , thus, affect the efficiency enough to transformation of E.coli. Moreover, water bath temperature may be not equilibrated (less than 42C or a higher which decrease in transformation efficiency ( Smith, et al, 1992).Also, the concentration of DNA has significant effect on the transformation efficiency , usually less amount of DNA is used. If using more, the result is fewer colonies because the impurities in the DNA will inhibit some of the cells from being transformed.There are main factors to consider during induction conditions Vector, Host Strain, and Growth Conditions. These thre e factors have the biggest impact on the expression of the protein of interest. First on the list of considerations is the vector that is used to express GFP protein. The first thing should be considered after cloning, the protein of interest is still in frame. It is recommended that before any experiment is carried out the first thing is should be done is cloned plasmid (or a few different clones) sequenced. This will show if the sequence you inserted into the expression vector is still correct and is still in frame. This is especially important if the construct contains any PCR fragments. If there are any point mutations or the sequence gets out of frame by even a few bases it can have dramatic effects on the protein that expressed. Another thing to check before expressing is if the GFP protein sequence contains long stretches of rare codons. This can cause the protein that is expressed to be truncated or non-functional. A few rare cordons spread around the protein are OK in most cases, but if there are a number of rare codons in a row, then it can have a big effect. The third sequence related step to optimize the protein production is to make sure there is not a high GC concentration at the 5 end of GFP protein. This could potentially cause problems with the mRNAs stability, and could prevent it from being translated correctly, which would also lead to truncated or non-functional proteins. If your sequence is GC heavy at this end, you can try to make a few silent mutations to break up long stretches to try and help stability.After the plasmid is sequence verified, the next factor is the bacterial host that is used. There are almost as may hosts as there are expression vectors, with certain hosts excelling in producing different types of proteins. For example if you have a toxic protein, or a protein that could potentially cause genomic rearrangement, you will want a vector that gives you very tight control over the induction of your protein. There can be le aky expression (i.e. expression of your protein without the addition of your inducer) that can potentially have adverse effects on the cells growth or even prevent your cells from over-expressing your protein in the first place. If youre utilizing the T7 polymerase system, then look for a host containing the pLysS plasmid, as this will code for T7 lysozyme, which will curtail the T7 polymerase and can greatly reduce the level of background expression. If as stated before you have a protein that contains a large number of rare codons, then look for a host with the genes for the necessary tRNAs already present, which should allow your protein to express correctly. Sometimes simply changing hosts can have a dramatic effect on the amount of protein produced and the stability of the protein that is made, so if one host isnt giving you the results you need, then feel free to switch your host up.The third and final factor to consider when expressing a protein is growth conditions. When fi rst starting out with the protein induction it is very important to run an expression time course, where you take a fresh colony from a streaked plate, and grow the culture to stationary phase. Next, dilute the overnight culture 1/100 and grow to mid log phase, then add the inducer and induce your protein for a number of hours, taking 1mL samples every hour or so. Once these samples are lysed, you can run an SDS-PAGE gel to determine your protein production levels. You might get great induction the first time, or you may have to tweak your conditions in order to get really good expression levels. Other factors that may need to be controlled for are the bacterial growth rate (determined by taking OD measurements during the induction process), and the temperature during induction. Some constructs will express perfectly fine at 37C, while others need to be bumped down to 30C to induce correctly. The concentration of the inducer too will have an effect, as many inducers (IPTG) can be to xic to the cells that they are inducing. Using freshly made inducer is good step to make sure you always have consistent results. Only through experimentation can you determine what will be best for your construct, and give you the most strong expression levels.Transformation efficiencyTransformation efficiency is a measure of the ability of cells to be transformed. Transformation efficiency is expressed as the number of transforms per mcg of pUC19.By using the following formulaColonies on plate / ng of control DNA X 1000ng/g = (transformation (T) / g plasmid DNA)100 L equivalent to 0.01 ng DNA in the plate.Growth curveIn general growth curve shows the S- shaped when plotted in log unidimensional format as shown in figure 4, that separated into four phasesLag phase the initial period when no increase in cell number is seen.Log phase when cells are growing at the maximumm rate.Stationary phase growth decreases as a nutrient are depleted and waste products accumulate. ending phase this is the result of prolonged starvation and toxicity.ConclusionThe main goal for the experiment was to express the protein of interest (GFP). However, factors influencing transformation efficiency include technique errors, the temperature and length of the incubation period, the growth stage of the cells, and using the correct mass of plasmid DNA. Escherichia coli is one of the most important hosts in modern day recombinant protein production. Throughout academia and industry its uses are widespread and with sequence data available for some of the most common strains of the bacteria it has been a favourite organism for many metabolic engineering and metabolic modelling projects in the past (Berry, 1996 Koffas et al., 1999).

The Morality Of Mercy Killing

The Morality Of Mercy KillingWhy is there such a affaire in our world that is called grace sidesplitting?, why? In this report I will discuss about euthanasia or tenderness extinguishing. euthanasia is the deliberate killing of a mortal for the benefit of that soul. In most cases euthanasia is carried out beca drop the person asks to die, but there are cases where a person cant make such a request. A person who undergoes euthanasia is ordinarily terminally ill. Euthanasia can be carried out either by doing something, such as administering a lethal injection, or by non kill a person even if they are terminally ill because this person needs a chance to live along and see his life, but there are people that disagree.IntroductionMercy killing, also known as mercy killing or assisted suicide is an act of murder which are usually to end the life of a person suffering from unadulterated pain or terminal illness. An individual who is tormented by a disease you want to plant at the end of his life, but do not have the ability to do so. In such a case, and may request assistance from an other person. This is il well-grounded in most, but not all, areas of the world. let certain languages, provided that they are to follow strict guidelines and the procedure is by a doctor, the patients request.In the United States, assisted suicide is legal in only ii states Oregon and Washington. And others have created, such as the Netherlands, Switzerland, Belgium and Luxembourg are also rules for legal mercy killing. Rules of praxis in areas where the law allows assisted suicide conscious.That regulations authorities the work of doctors allowed to put an end to the suffering of patients who choose voluntarily to physician assisted suicide. The case of terminally ill, which creates severe physical pain may await assistance. In areas where it is permitted by the physician assisted suicide is generally not legal for any person other than a doctor to help in the death of the last, however.Mercy killing and assisted suicide is a controversial issue in the community and among medical professionals in general. Some doctors believe that the practice of assisted suicide is incompatible with the teaching profession does not hurt. On the other hand, others argue that extending the life of a person who wishes to die is a impact of human rights harsh.One of the most popular activities assisted suicide is an American pathologist Dr. Jack Kevorkian. She helped illegally in the death of more than than 100 patients in the United States, was convicted of second-degree murder in late 1990. In 2007, he was released from prison. Since then, Dr. Kevorkian appeared in several universities in the talk shows to promote his beliefs and theories about the patients right to die.While it is more common, and not all of mercy killing with the help of a doctor. Although it is a criminal act, there are many reported cases in one family portion or other non-medical staff unti l the life of another, usually at the request of the patient. In some cases this is done by removing or cutting the life-saving art. At other times, the act was committed by administering a lethal overdose of drugs. If convicted, the person who can send mercy killing is illegal to prisonClassification of mercy killingAnd mercy killing can be classified according to whether the person giving informed consent in three types voluntary and non-voluntary and involuntary. in that location is controversy in the medical literature and bioethics about what if it can be considered as non-voluntary (and by extension, it was accidental) killing patients, mercy killing, regardless of the intentions or the circumstances of the patient. In the definitions offered by Beauchamp and Davidson, and then by the Wreen, and consent by the patient which are not considered to be one of the criteria, although it was asked to justify mercy killing. However, others see the agreement as necessary.Voluntary mer cy killingMercy killing was carried out with the consent of the patients so-called voluntary mercy killing. And active voluntary mercy killing is legal in Belgium, Luxembourg and the Netherlands. Voluntary passive mercy killing is legal in the United States Cruzan V. Director of the Department of Health and the state of Missouri. Is often utilize when the patient brings about his death for the assistance of physician-assisted suicide instead. Suicide is legal in Switzerland and the United States from Oregon, Washington and Montana.Non-voluntary mercy killingThe implementation of mercy killing if the patients consent is not available so-called non-voluntary mercy killing. For example, mercy killing, which is illegal in all parts of the world, but in the criminalization of certain precise circumstances in the Netherlands with the Groningen Protocol.Involuntary mercy killingPerformed mercy killing against the wishes of the patients so-called non-voluntary mercy killing.Procedural de cisionCan be divided into voluntary and non-voluntary mercy killing and involuntary to more variables, passive or active. A number of authors to consider these conditions to be misleading and unhelpful. nonoperational mercy killingPassive mercy killing involves the rejection of common treatments such as antibiotics, which are necessary to sustain life.Last mercy killingLast mercy killing involves the use of lethal substances or forces to kill the most controversial.Sense of medicalThe poll comes in the United States more than 10000 doctors to the conclusion that it was thought about 16% of the doctors to stop life-sustaining treatment because the familys request, even if he believed it was too premature. About 55% do not, and the rest 29% depends on the circumstances.This study also far-famed that California. 46% of doctors agree that he should be allowed to physician-assisted suicide in some cases 41%, no, and the remaining 14% think it dependsDiscussionEven some who stand and we can say the above principles, there is no real difference between allowing someone to die and the status of the person suffering in a coma until his death, in order to alleviate the suffering. Then you have to choose not to remove life-support establishments. After all, as they say, and the result is the same in both cases. Also, if you remove your old age or treatment of a patient dying, the patient dies because of this procedure. Thus, they contend, by removing life-prolonging treatment is. A case of murder in the mercy killing as the cause of death is the direct intervention by the person injecting the performance or act of violence after death. Although the ii procedures, euthanasia, and allowing to die, is similar in the result, are not the same in process or in the grounds next.In the case of allowing to die, it is true that the patient usually dies on the removal mechanisms of aging or treatment, but the cause of death is a condition that there is now allowed to have a nat ural effect. For example, the device is removed the ventilator and the patient died because of disease, cardiovascular system and lungs. Mechanism or treatment that prevent the removal of this life-threatening disease because it is more useful cognitive function and emotional, are not allowed to be a disease that affects the natural. Allows nature to take its course. In euthanasia, and the cause of death is a disease caused by a mercy killer. Can cause disease by direct interference with the normal activity of the physiological system of the patient or to withhold specific treatment for the system that should be provided. Can therefore be achieved by withholding the necessary euthanasia drug, as well as a pistol. In one way or another, therefore, euthanasia is an act of violence committed in the physiology of the person concerned and which caused the disease. In addition, at least an implicit apology of euthanasia is to exercise total domination over the human life.Some argue that t he ultimate justification for euthanasia, and to liberate the suffering, is enough to justify this action. In this way, as they say, Roswell Gilbert, and others who wish to put an end to the suffering should not lead to the imposition of moral or legal. Unfortunately, there are many ethicists today to justify any action until the last terra firma is good. But such thinking ignores the reality. Actions that lead to achievement motivation and one that will eventually be a reason to be morally justified. For example, if the final was my motivation may be to collect money to send my children to university, and I do not have the right to abstract the money of widows and orphans.ConclusionMercy killing can and do not go hand in hand.According to Buddhism, can not justify mercy killing. Mercy killing can and do not go hand in hand. Some people kill pets because they do not like seeing animals suffer.Some try to justify mercy killing with the misconception that if the penury or reason f or this is good, then the act itself is a good thing. Then that the claim to kill pets, they have the intention to alleviate the suffering of the unfortunate animal, and so a good job. There is no doubt that the original intent or motive is good. But the act of killing the evil that is done during the second idea would certainly help in achieving health outcomes.final thought To determine the measure out or not intended in any business and we realize that may be the person who conducted the work of a self-satisfied with any sense of moral guilt. In the case of Roswell Gilbert and all the others in this same position, and we will be more interested in the discussion of the support system that would allow him to achieve his pain more humane way to discuss what could be a just punishment.

Japan Recession Essay examples -- essays papers

Japan Recession Thomas Paine was born on January 29, 1737 at Thetford, Norfolk in England, as a son of a Quaker. In 1774, he met Benjamin Franklin in London, who advised him to immigrate to America, giving him letters of recommendation. Paine reached Philadelphia on November 30, 1774 where he started over as a publicist. He wrote many articles that fey on various topics. On January 10, 1776 Thomas Paine grouped his ideas on American independence in his pamphlet Common Sense. I think Thomas Paines Common Sense had a significant influence on the separation from England. I believe Paine thought the Colonies had all the right to revolt against a presidential term that imposed taxes on them but didnt give them the right of representation in the Parliament at Westminster. Thomas believed there was no reason for the Colonies to incumbrance dependent on England. I think Thomas Paine had an incredible way of persuading people to take action through his writing. In his Common Sense, Paine states that sooner or later independence from England must come, because America had lost touch with the mother country. In his words, all the arguments for separation of England are based on nothing more than simple facts, plain arguments and common sense. Paine viewed government as necessary evil that could only become safe when it was represented and altered by frequent elections. Paine to a fault states that the function of government in society ought to be only regulating and as simple as...

To Dam, or Not To Dam Essay -- Marc Reisner Ecology Environmental Essa

To Dam, or Not To DamThe River, slightly milky from glacial sediment, tumbles down rocky chutes, boils through tight canyons, and glides across beds of agatelike stones. In the distance, poking through storm clouds, ar plunging slopes dense with virgin hemlock and firIntruding into this primeval scene are two faint dams (Reisner, 382). In this essay by Marc Reisner, his standpoint on the issue of dams is very well seen. Reisner talks of the ecological damages that dams create. The debate oer dams has been heated in recent years, their harmful effects and overall abundance is the topic for such debates. But are dams as bad as everyone says they are, do dams do more(prenominal) harm than good, or more good than harm? Emphasis should be placed on comparing the ecological effects verses the economical benefits. There are many pros and cons for the ecological side of this debate. One pro is that dams help areas that would otherwise be waterless and barren support life. Taken from a pamphlet hustling by the deputation on Public Awareness and Education, Water is the vital resource to support all forms of life on earth. Unfortunately, it is not evenly distributed over the world by season or location. Some parts of the world are prone to drought making water a scarce and incomparable commodity, while in other parts of the world it appears in raging torrents causing floods and loss of life and property. Throughout the history of the world, dams and reservoirs have been successfully in collecting, storing and managing water needed to sustain civilization (CPAE, 1). This is a major benefit to the ecology of the world, as the quote stated water is the basis of life, the human remains depends on water for survival. Although ther... ...to benefit the United States. Even those who oppose dams benefit from them in one way or another, destroying them would give our nations economic system a detrimental blow. Finally for the solution for this dilemma, I dont believe on that point is an general solution. Each dam should be looked at individually, if the costs outweigh the benefits, then it should be destroyed, and vice versa. The issue of dams will not be easily solved. Ever since the first dam was built there has been controversy, and as long as one stands so will the debate. ReferencesMcKibben, Bill. Daybreak. In A Forest of Voices Conversations in Ecology 2nd Edition (pp. 156-169). Mayfield Publishing Company.Committee on Public Awareness and Education. (1997, May). Benefits and Concerns About Dams an Argumentaire. Retrieved March 3, 2003, from http//www.icold-cigb.org/BandC.PDF