Les2_ppt_blackwhite

Les2_ppt_blackwhite - Chapter II Reporter systems and...

Info iconThis preview shows pages 1–8. Sign up to view the full content.

View Full Document Right Arrow Icon
Chapter II: Reporter systems and detection methods
Background image of page 1

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

View Full Document Right Arrow Icon
Outline 1. What? Why? 2. Basic principles 3. Common reporter systems: 1. What, detection 2. Applications, recent developments 3. Advantages/disadvantages
Background image of page 2
1. What? A reporter gene = gene whose phenotypic expression is easy to monitor/visualize
Background image of page 3

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

View Full Document Right Arrow Icon
Why were reporter systems developed? • Protein of interest: – No biochemical activity – No enzymatic activity – Hard to visualize – Lack of good antibodies – …. . • Companies: HT assays
Background image of page 4
To control. .. To control. .. – genetic construction process genetic construction process (blue/white screen) (blue/white screen) – transfection transfection processes and efficiency processes and efficiency To detect gene expression and regulation To detect gene expression and regulation – Promotor Promotor /enhancer activity /enhancer activity – Regulatory events Regulatory events ( e.g. effect of inhibitors on promotor activity, to determine essential TF-binding sites within a promotor,…) – Recombination events ( Recombination events ( Cre Cre /Lox /Lox ) To visualize cellular processes To visualize cellular processes (( (( trans)location trans)location , PPI, signaling, , PPI, signaling, …) Why use reporter systems? Hey! I’ve been produced by this cell. ..
Background image of page 5

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

View Full Document Right Arrow Icon
2. Basic principles/requirements • Cheap, easy and fast (HT!!) • Reporter half-life • Reporter gene (activity) quantification and visualization: lum, fl, RA, color, WB, ELISA
Background image of page 6
• Reporter: – Reporter could be detected Reporter could be detected as such as such without without substrate substrate addition addition (e.g. fluorescent proteins) (e.g. fluorescent proteins) – Reporter = Reporter = enzyme enzyme : detected by substrate conversion : detected by substrate conversion
Background image of page 7

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

View Full Document Right Arrow Icon
Image of page 8
This is the end of the preview. Sign up to access the rest of the document.

{[ snackBarMessage ]}

Page1 / 39

Les2_ppt_blackwhite - Chapter II Reporter systems and...

This preview shows document pages 1 - 8. Sign up to view the full document.

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