|Register||Search||Today's Posts||Mark Forums Read|
|DNA Techniques Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.|
| ||LinkBack||Thread Tools||Display Modes|
EGFP Translation Study Problem
I am performing a translation study in mammalian cell culture. My construct includes: universal promoter of the vector, leader sequence of my gene of interest (which has a meaning in a translation efficiency), EGFP and FLAG tag. Using antibodies against FLAg I am able to obtain band of fusion protein of expected size in western blot - therefore I concluded that translation takes place in my culture. But EGFP signal is absent, when cells are observed under the fluorescent microscope.
Any ideas why I am having this problem?
Re: EGFP Translation Study Problem
based on ur observation it is highly possible that your fusion protein (GeneX-EGFP-FLAG) does not fluoresce. EGFP is quite sensitive, any excess amount of residues fused to it will drastically alter its folding properties, hence its fluorescence intensity.
|egfp , problem , study , translation|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Human Cytome Project - Update 24 Jan. 2005||Peter Van Osta||Cell Biology and Cell Culture||1||08-01-2010 02:18 PM|
|Cecità congenita e sogno||Aleph||Forum Biologia||53||11-06-2006 02:03 PM|
|Human Cytome Project - How to explore - Update 19 April 2005||Peter Van Osta||Cell Biology and Cell Culture||0||04-19-2005 07:10 AM|
|Human Cytome Project - Update 6 Jan. 2005||Peter Van Osta||Cell Biology and Cell Culture||0||01-06-2005 10:18 AM|
|Druc discovery, diseases and the Human Cytome||Peter Van Osta||Cell Biology and Cell Culture||0||09-24-2004 07:42 AM|