MICB 325 Study Guide - Quiz Guide: Mhc Class I Polypeptide-Related Sequence B, Start Codon, Restriction Site
Document Summary
Perhaps there is a regulatory region, as well as a promoter, between the crt atg start codon and the bamh i site. Use of a reporter" coding sequence to measure crt promoter activity. We can use the lacz coding sequence of e. coli because r. capsulatus does not make a -galactosidase. In this particular case we could use the crt gene and measure carotenoid synthesis as an index of promoter activity, but instead we will use a lacz reporter gene to illustrate the basic principle. Various reporter genes exist: use pcr to amplify segments of crt promoter region; moving in from the bam end, ligate shortened segments 5" (transcriptionally upstream") of a promoterless lacz coding sequence on a plasmid. Note that we can add a restriction site to the pcr product, either internally or as a tail on 5" end of pcr primer (explained in lecture), to allow subsequent cutting and ligation.
