Spring 09 lecture 21

Spring 09 lecture 21 - Start site of transcription Start...

Info iconThis preview shows pages 1–6. Sign up to view the full content.

View Full Document Right Arrow Icon
Run-on, run-off and reporter gene transcription (methods)& mRNA is getting transcribed during the assay W information about the actual efficiency of transcription – transcription rate These assays are used to locate AND assay the promoter region ( presence/absence of all necessary elements, their position and availability ) Start site of transcription Upstream cis regulatory elements located in this region (= promoter) Transcribed region Downstream cis regulatory elements may be located in this region Start site of translation Stop site of translation GENE
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
rrently the most frequently used are reporter genes de for easily assayed gene products (usually enzymes): Idea: to investigate the strength of the promoter you clone the reporter gene after promoter of interest u expression of the reporter gene is under control of promoter of interest u 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 Will give you info on how important the deleted part is
Background image of page 2
http://www.forbes.com/2001/07/26/0726gfp_print.htm Lac Z product β- galactosidase - hydrolyses X-Gal (substrate) into a colorless galactose and blue 4-chloro-3-brom-indigo CAT - c hloramphenicol a cetyl t ransferase, acetylates antibiotic chloramphenicol (CAM) & acetylated CAM has different properties then un-acetylated Luciferase - ATP and luciferin (substrate) & emission of light GFP - green fluorescent protein from bioluminescent jellyfish Aequoria Victoria (naturally fluoresces green when exposed to blue light – not an enzyme) Reporter genes - easily assayed gene products
Background image of page 3

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
RNAPII promoters Learning Objectives Describe the cis -elements commonly associated with eukaryote genes as well as RNAPII promoter specific elements Define what is meant by the modular nature of eukaryote promoters
Background image of page 4
(Lewin, 2006. Fig. 24.1) promoter: operationally defined (very simplified definition): those sequences that must be located in the vicinity of the start point & are required for initiation of transcription In most cases class II promoters (promoter regions) have two parts: 1.
Background image of page 5

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Image of page 6
This is the end of the preview. Sign up to access the rest of the document.

This note was uploaded on 08/03/2009 for the course BIOL BIOL308 taught by Professor Miskovic during the Spring '09 term at Waterloo.

Page1 / 22

Spring 09 lecture 21 - Start site of transcription Start...

This preview shows document pages 1 - 6. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online