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| DNA Techniques Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins. |
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#1
01-14-2009, 11:49 PM
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 EGFP Translation Study Problem Hi all! 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? Thanks  |
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#2
10-14-2011, 04:23 PM
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| 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. |
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| egfp , problem , study , translation |
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