Posted by
Michael Schell on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Flexible-linkers-tp5600224p5600431.html
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We've mostly used GGSGS, which is flexible with a bit of hydrophililc character. We did a comparison whereby we compared this linker to no linker whatsoever. The EGFP fluorescence was far brighter with the linker, presumably because the EGFP folded better.
However, there have been situations where a stiff or bulky linker (DPPVAT, the inker formed when cloning into EGFPN1; or YSDLELKF from pEGFPC3) turned out to be preferable. We assume that, in these cases, the kinks created by the dual prolines or else aromatic side groups push the GFP away from the protein it is fused to, and minimizes some sort of undesirable steric hindrance. In lieu of structural info, a certain amount of guesswork is unavoidable.
Mike
Michael J. Schell, Ph.D., CIV, USUHS
Assist. Professor
Dept. of Pharmacology
Uniformed Services University
4301 Jones Bridge Rd.
Bethesda, MD 20814-3220
tel: (301) 295-3249
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>>> Sean Speese <
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This is a tad bit off topic, but I was wondering if people would be
willing to share their experience with flexible linkers used when making
fusions of your protein of interest to fluorescent proteins. What are some
sequences that seem to work well? This will likely be empirical, but still
useful information for me in deciding what linkers to use in the future.
Thanks,
Sean Speese
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