Biochemistry 2280A Lecture Notes - Lecture 27: Recombinant Dna, Escherichia Coli, Cloning
12 Jun 2016
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You have identified a human protein that will be of tremendous value in treating disease (e. g. hgh) Your goal is to mass produce this protein. The process of expressing yfp will require: cloning yfg, introducing yfg into e. coli, purifying the protein from e. coli. Why are bacteria the organism of choice for expressing yfg: they grow quickly and inexpensively, protein extracts are made easily (easy to break bacteria and release the protein. Bacteria cell walls are fairly rigid but not as strong as other organisms: genetic engineering is simple (bacteria have been used in genetic engineering for 40 years, multicopy plasmids and strong promoters can drive expression. 5% of the total bacterial protein may be your protein! First issue: we need a source of the gene. Only for a few rare genes that lack introns: less than 10% of human genes lack introns, some organisms like yeast have very few introns so we can use them.
