01:694:215 Lecture Notes - Lecture 7: Reverse Transcription Polymerase Chain Reaction, Northern Blot, Ribosomal Rna

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To identify the proteins at the identified sites (mutations) Screen for mutants in genes that are required for meiosis and sporulation. You don"t want to screen for plasmids with the mutant because we already know there is a mutation at the site. Only the cells with the mutated proteins will be white. But you do not have to worry about that because. Plasmid have many copies in the yeast. The chance of all of them picking up a mutation is very small. Thus, the mutation is likely due to the protein. Heterologous- introduce wild type into different promoter. Even some lacz activity would make the cells appear blue. Therefore, all plasmids need to pick up mutant in order for failure to occur (white) On the other hand, for a repressor site, the wild type would originally be white, but one mutant plasmid is enough to turn it blue. Find the main activator/repressor driving the gene expression/activity.

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