BIOL308 Lecture Notes - Rna Polymerase Ii, Green Fluorescent Protein, Chloramphenicol
16 Jul 2014
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Document Summary
Currently the most frequently used are reporter genes. Code for easily assayed gene products (usually enzymes): Idea: to investigate the strength of the promoter, you clone the reporter gene after promoter of interest. Expression of the reporter gene is under control of promoter of interest. Level of expression of measurable (easy to assay) gene product (enzyme) is proportional to the strength of the promoter. You can delete/mutate a part of the promoter starting from 5" end (5" deletion series) and repeat the experiment. Build a map of the control region with indications of relative importance of sequence in that area. Lac z product, beta-galactosidase; hydrolyses x-gal (substrate) into a colorless galactose and blue 4- chloro-3-brom-indigo. Cat; chloramphenicol acetyl transferase, acetylates antibiotic chloramphenicol (cam) Gfp; green fluorescent protein from bioluminescent jellyfish aequoria victoria. Naturally fluoresces green when exposed to blue light; not an enzyme. Promoters recognized by rnap ii - class ii promoters.
