 ideas for Fluorescent tagging of 7Kda bacterial protein. |
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  Nov 2 2009, 07:42 AM
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member  Group: Members Posts: 1 Joined: 2-November 09 Member No.: 13692  |
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? 
This post has been edited by Afroscience: Nov 2 2009, 07:43 AM |
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Nov 3 2009, 03:41 PM
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#2
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member Group: Active Members Posts: 9 Joined: 24-September 09 Member No.: 12709 |
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!
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