BIOL 200 Lecture Notes - Lecture 32: Two-Dimensional Gel Electrophoresis, Dna-Binding Domain, Reverse Genetics
Document Summary
Reverse genetics: know sequence, don"t know function: ex. Know sequence - make dsrna that correspond to the sequence, introduce it and then look at phenotypes. Forward genetics: working on function and go back and clone the gene/ nd the sequence. Can assess protein modi cations via 2d gel electrophoresis: identify polypeptides within the cores using mass spec - will tell what proteins are in original mixture by comparing to a data base. Can understand more about protein networks using yeast. Transcription factors are modular: they consist of a dna binding domain that will recognize speci c sequences, coupled to a transcription activation domain - required to activate transcription, can mix and match modules. Ie take binding domain from gal4 and stick it to any transcriptional activation domain. Hybrid transcription factor will activate transcription in a uas(upstream activation sequence)-dependent manner: will summon in transcription factors and turn on transcription of downstream gene, can mix and match and transcription factors work!
