CMMB 549 Lecture Notes - Lecture 22: Transposable Element, How Do You Know, Plasmid
28 Dec 2019
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Do hybridizations using plasmid gene as probe. Limitation of mutagenesis: to make mutants using forward genetics approach, without a good screen to identify mutants, you have problem. 2 methods designed to get around limitations of mutagenesis. Some assay work with broad spectrum pathogens which are not real pathogens at all in some cases. Basic principle: make pools of mutants, one from each sequenced tagged set of mutants. Inoculate into animal, at end of experiment look for sequence tags that are missing from mutant. You have a lot of versions of transposon, each with different sequence tag in it. Sometimes tags are different lengths, sometimes need to hybridize to identify them. Inserted into transposon is an amplifiable piece of dna that is different in each transposon. Do transposon mutagenesis experiment with each transposon, and you get separate pools of bacteria mutagenized with each different sequence tag transposons.
