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Name: sid
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Question:
Can you explain Insulin production in bacteria and its regulation by IPTG.



Replies:
Information on expression of eukaryotic genes in bacteria can be found in any molecular biology textbook. See for instance 'Recombinant DNA by Watson, Gilman, Witkowski and Zoller, 2nd Ed., chapter 23 where the cloning of insulin is described. The use of an IPTG inducable promoter (the promoter of LacZ is only active when the inductor IPTG is present) is also explained in text books, and even in catalogs of the companies selling the plasmids that are used for such experiments. With a bit of reading you'll become an expert!

Dr. Wassenaar


I'm not sure what IPTG is. But are you referring to recombinant DNA technology? First a human gene for insulin was isolated and cut out of the human chromosome with restriction enzymes. These are enzymes that cut DNA at very specific spots in the DNA. They are like DNA scissors. Then a small piece of DNA called a plasmid is isolated from a bacteria. The same restriction enzyme is used to cut the plasmid. The insulin gene from the human is inserted into the bacterial DNA and they are sealed together with an enzyme called ligase. The plasmid is reinserted into the bacteria and the bacteria will treat the human insulin gene as its own. When it comes time to make protein it will make the insulin as well. Bacteria reproduce very rapidly and are easily maintained. We can get vats of human insulin by this method.

Van Hoeck



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