Rdna Technology

A) What specific biotechnology application did you choose? Recombinant DNA technology is used in producing insulin from E.coli Insulin is expressed in recombinant E. coli and then subsequently purified and refolded in to bioactive form. Cloning of insulin, expression in different host, fermentation process optimization, refolding and purification of active protein from inclusion bodies and finally formulation of insulin having better pharmacokinetics has provided model system for production of most of the therapeutics using r-DNA technology. B.Briefly describe this biotechnology application. Insulin used for diabetes was earlier extracted from pancreas of slaughtered cattle and pigs. Insulin from an animal source, though caused some patients to develop allergy or other types of reactions to the foreign protein. Insulin consists of two short polypeptide chains: chain A and chain B, that are linked together by disulphide bridges. In mammals, including humans, insulin is synthesised as a pro-hormone (like a pro-enzyme, the pro-hormone also needs to be processed before it becomes a fully mature and functional hormone) which contains an extra stretch called the C peptide. This C peptide is not present in the mature insulin and is removed during maturation into insulin.The main challenge for production of insulin using rDNA techniques was getting insulin assembled into a mature form. In 1983, Eli Lilly an American company prepared two DNA sequences corresponding to A and B, chains of human insulin and introduced them in plasmids of E. coli to produce insulin chains. Chains A and B were produced separately, extracted and combined by creating disulfide bonds to form human insulin. This application is explained in seven steps Gene Isolation Preparing Target DNA - Inserting DNA into Plasmid - Inserting Plasmid back into cell Plasmid multiply Target Cells Reproduce Cells Produce Proteins Step 1. Gene Isolation The gene for HUMAN insulin producing protein is isolated. The gene is part of the DNA in a human chromosome. The isolated gene is then copied so that many insulin genes are produced. Step 2. Preparing Target DNA Recombinant DNA technology is the process of taking DNA from one organism and put it in the DNA of bacterium. From a bacterial cell a circular piece of DNA called a plasmid is removed. for cutting the plasmid ring open Special proteins are used . Setp 3. Insert DNA into Plasmid The gene for insulin is inserted into the opening end of the plasmid ring and the ring is closed. The human insulin gene is now recombined with the bacterial DNA plasmid. Step 4. Inserting Plasmid back into cell The bacterial DNA now contains the human insulin gene and is inserted into a bacteria. Scientists use very small needle syringes to move the recombined plasmid through the bacterial cell membrane. 5. Plasmid multiply Many plasmids with the insulin gene are inserted into many bacterial cells. The cells need nutrients in order to grow, divide, and live. While they live, the bacterial cell processes turn on the gene for human insulin and the insulin is produced in the cell. When the bacterial cells reproduce by dividing, the human insulin gene is also reproduced in the newly created cells. 6. Target Cells Reproduce Human insulin protein molecules produced by bacteria are gathered and purified. The process of purifying and producing cow and pig insulin has been greatly reduced or eliminated. 7. Cells Produce Proteins Millions of people with diabetes now take human insulin produced by bacteria or yeast (biosynthetic insulin) that is genetically compatible with their bodies, just like the perfect insulin produced naturally in your body.

B) Based upon the definition of biotechnology, describe how your biotechnology application is an example of biotechnology. The term biotechnology is a fusion of biology and technology Biotechnology refers to the engineering of phenotype of an organism. DNA technology is the chemical manipulation of the genotypes and resulting phenotypes of organisms such that living organisms are modified. Both are modifying the phenotype of an organism. So we can consider rDNA technology is an example of Biotechnology

C) What benefits are derived from this application? A special committee of the National Academy of Sciences specifically reviewed the issues on the introduction into the environment of organisms genetically engineered using recombinant DNA technology. They concluded that "there is no evidence that unique hazards exist either in the use of R-DNA technique or in the transfer of genes between unrelated organisms," and that "the risks associated with the introduction of R-DNA engineered organisms are the same kind as those associated with the introduction of unmodified organisms." The committee concluded that rDNA techniques constitute a powerful and safe new means for the modification of organisms for the benefit of animals and humans. They also stated that there is adequate scientific knowledge to guide the safe and prudent use of such organisms outside research laboratories.

Advantages of Recombinant technology: .Provide substantial quantity .No need for natural or organic factors .Tailor made product that you can control .Unlimited utilizations .Cheap .Resistant to natural inhibitors

D) Are there any concerns or negative impacts associated with this use of biotechnology? Why or why not? Irrespective of the expression system used for the production of recombinant molecules, all biotherapeutics share major safety concern. This is due to inherent nature of the biological molecules, where the conformation of the molecule is more important along with the chemical nature. Thus, any biomolecules manufactured using a living system need complete characterization and safety evaluation. The most important criteria during the manufacturing of recombinant molecules using live organism are the quality, safety and efficacy. All these three parameters influence the toxicity and immunogenicity of the final product and thus needs critical evaluation. some of the parameters associated with different types of product. It is most essential to verify this parameter so that purity, safety and efficacy is maintained not only during production but also during storage and application. It is also essential that all precautions are taken at each step of biological production taking care of the biosaftey aspects. The host organism and waste product coming out of the r-DNA process need to be processed according to the environmental protection act of the country so that biodiversity is not affected Some of the risk associated with Recombinant DNA (r DNA) proteins are Quality, stability, Species specificity, vector and transgene toxicity, including immunogenicity and species relevance for activity, toxicity related to the pharmcodynamic effects, biological toxicity, local tolerance, onco genicity

E) would you happen to know anything on embryonic stem cells? Embryonic stem cells are derived from embryos. Most embryonic stem cells are derived from embryos that develop from eggs that have been fertilized in vitro-in an in vitro fertilization clinic-and then donated for research purposes with informed consent of the donors. They are not derived from eggs fertilized in a woman`s body