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[Afroscience]

Hi am attempting to localize a small 7kDa DNA binding protein in E. coli cells via fluorescent microscopy. ....GFP and his patches on c or n terminus kills function.. I am thinking of using either genetically encoded florescent amino acid   on the back  of the protein or using a His-patch on the protein and then use anti-His fluorescent anitbodies to localize the protein in vivo....  also under consideration was the use of tetra cysteine motifs...
Does any one have any suggestions or advice concerning the methods i am contemplating?

Edited by Afroscience, 02 November 2009 - 07:43 AM.

[Holsten]

You can try fuse it with small tags like V5 or HA, hopefully they wouldn't kill the function of your protein. GFP is not a good choice in your case - it's too big for your protein and tends to form dimers. Good luck!