BIOL 200 Lecture 31: Lecture 31 - Functional Genomics & Proteomics
Document Summary
Make a vector with anking t7 rna pol promoters that drive transcription of a region between them that investigators inserted a sequence that corresponds to every single gene in the genome. Transform 19,000 times so that you have 19,000 bacterial colonies each of which can expressed a dsrna of a given gene in the genome. A variety of genes were found to give rise to the various visible phenotypes. Genes that cause embryonic lethality and sterility were often involved in basal cellular function (essential genes; ie translation, transcription, splicing) The disruption construct is introduced into diploid yeast cells to replace the appropriate region: replace region with a selectable marker - mediated by homologous recombination. Flanking sequence has to be 100% homologous to. Presence of the dominant selectable marker confers drug resistance (g-418) so the cells can grow on drug: drug resistance gene has been incorporated into a strand in the chromosome.
