ideas for Fluorescent tagging of 7Kda bacterial protein. - (Nov/02/2009 )
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?
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!