{[ promptMessage ]}

Bookmark it

{[ promptMessage ]}

Spring 09 lecture 21

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

Info icon This 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
Image of page 1

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

View Full Document Right 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
Image of page 2
http://www.forbes.com/2001/07/26/0726gfp_print.htm l 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
Image of page 3

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

View Full Document Right 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
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.
Image of page 5

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

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

{[ snackBarMessage ]}

What students are saying

  • Left Quote Icon

    As a current student on this bumpy collegiate pathway, I stumbled upon Course Hero, where I can find study resources for nearly all my courses, get online help from tutors 24/7, and even share my old projects, papers, and lecture notes with other students.

    Student Picture

    Kiran Temple University Fox School of Business ‘17, Course Hero Intern

  • Left Quote Icon

    I cannot even describe how much Course Hero helped me this summer. It’s truly become something I can always rely on and help me. In the end, I was not only able to survive summer classes, but I was able to thrive thanks to Course Hero.

    Student Picture

    Dana University of Pennsylvania ‘17, Course Hero Intern

  • Left Quote Icon

    The ability to access any university’s resources through Course Hero proved invaluable in my case. I was behind on Tulane coursework and actually used UCLA’s materials to help me move forward and get everything together on time.

    Student Picture

    Jill Tulane University ‘16, Course Hero Intern