Biochemistry 2280A Lecture Notes - Lecture 21: Northern Blot, Southern Blot, Hybridization Probe
30 Dec 2017
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You have identified a human protein that will be of tremendous value (ex. Human factor viii, hgh, insulin, interleukins: goal: to mass produce this protein using recombinant dna techniques. Why is expression of yfp using recombinant dna tech important: many proteins of or commercial or research use are very difficult to obtain from their native source, rdt can dramatically increase expression and facilitate purification. Process of expressing yfp requires: cloning yfg, purifying the protein from e. coli. E. coli are often used to express yfp: grows quickly and inexpensively, genetic engineering is simple, multicopy plasmids and strong promoters can drive expression, protein extracts are easily made. Introns are a problem in expressing eukaryotic genomic dna in e. coli: use mrna (cdna) to clone yfg, mrna does not contain introns (gets spliced out, each mrna encodes one gene. Finding a tissue that expresses a lot of yfg: nucleic acid hybridization:
