Microbiology and Immunology 2500A/B Lecture Notes - Lecture 33: Gene Knockout, Escherichia Coli, Dna Sequencing

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Novel methods have been devised to obtain large amounts of dna sequence at minimal cost. Rna pro ling (rna-seq) - instead of hybridizing, sequencing all the rna in a cell not just choosing ones you want. E. coli are the most commonly used organism for expressing yfp. Some things to consider when expressing your favourite human protein in e. coli. E. coli does not have the same modi cation mechanisms that our cells do. Bacteria probably will not work for this cause. Human & e. coli promoter structure being different, and different mechanisms of translational initiation are problems that are handled by the use of plasmids for cloning. Insert favourite gene after e. coli promoter sequence which is already contained on the plasmid so that when you insert the plasmid into the e. coli, it will already be programmed by the plasmid to work. Must have correct e. coli promoter sequence and ribosome binding site.

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