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Recombinant DNA technology used for insulin production

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Introduction

It was not until 1921 that diabetes was assumed to have been caused by a malfunction in the digestive system which was known to be related to the pancreas gland, however it was discovered that the disease was caused by the deficiency of insulin. At first, scientists put effort in synthetically recreating the insulin protein by putting together the sequence of amino acids in a chemically; however, they struggled to produce it in high amounts. Therefore, the insulin hormone was obtained by purifying it from animals, primarily cattle and pigs. Nevertheless, it caused side effects on people ?such as skin rashes- because the insulin purified from animals didn?t exactly match with the human insulin there are one or three amino acids that differ human insulin from animal insulin. In 1978, with the help of biotechnology, the human insulin gene was inserted into bacterial DNA and used to produce the insulin in large amounts. ...read more.

Middle

Eco RI recognises the sequence ?GAATTC? on the forward strand and ?CTTAAG? on the parallel strand of the DNA, forming staggered cuts between the G (Guanine) and the A (Adenine) on both strands. Using PCR (Polymerase Chain Reaction), the isolated human insulin gene is copied in order to allow a very large number of genes available to work with. For confirming the sizes (how many base pairs long the gene sequence is) of the insulin genes that were copied, a gel is run as Gel Electrophoresis is performed. The DNA ladder being used identifies the size of the insulin gene being 1430 base pairs long. (Genetic Home Reference, 2015) These staggered cuts leave sticky ends, which can form hydrogen bonds with the complementary sequence on the E.coli plasmids (circular DNA found in bacteria) which are also cut using the Eco RI restriction enzyme once it is removed from the bacteria cell. ...read more.

Conclusion

This step is called, Bacterial Transformation. Nutrient is given to the bacteria cell to maintain its function; to divide and live. Meanwhile they produce the human insulin, as the bacterial cells function and activate the human insulin gene. Therefore, as the bacteria cells grow and divide, the human insulin gene is passed down to the newly formed cells, allowing the insulin to produce furthermore. With the fermentation process, large quantities of bacteria are produced, and when it is reached to a sufficient amount, the bacteria containing the recombinant DNA are removed from the fermentation tanks in order to gather and purify the human insulin protein molecules the bacteria produce. The bacteria cells are destroyed once the process comes to an end. Nowadays, people with diabetes acquire human insulin from bacterial sources which is advantageously compatible with their own bodies, not at all different from the insulin produced naturally by humans. (778 words) ...read more.

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