Biochemistry 3380G Study Guide - Final Guide: Prokaryote, Ethidium Bromide, Ribonuclease H
8 Feb 2015
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Topic 26 recombinant dna technology: recombinant dna is possible because dna molecules from all organisms share the same chemical structure they only differ in the nucleotide sequences. Case study: you have identified a human protein that will be of tremendous value in treating disease (eg. hgh, your goal is to mass produce this protein. Why is expression of your favourite protein using recombinant dna technology. Important: many proteins are difficult to obtain from their native source, rdt can dramatically increase expression and facilitate purification. The process of expressing yfp require: cloning yfg. Answer: only for a few rare genes that lack introns. Introns are a problem in expressing eukaryotic genes in e. coli. We will use mrna (cdna) to clone yfg: mrna/cdna by definition does not contain introns, each mrna/cdna encodes one gene. First step is to determine which tissues express yfg done through hybridization/northern blot. Nucleic acid hybridization: denaturation / melting, renaturation / annealing / hybridization.
