3 replies to this topic

[Pangea]

Lets say İ would like to clone my Gene BamHI and Not1 into pGEX GST N. When İ do that i will get additional Amino acids on my rProtein after cleavge with thrombin. İ think gly will not effect my overall structure but nevertheless its there.

[bob1]

It is a common problem, the only real solution is to PCR a short tag (e.g. FLAG, HA) onto your DNA sequence and use that instead of the GFP

[Pangea]

bob1, on 03 January 2013 - 02:06 PM, said:

It is a common problem, the only real solution is to PCR a short tag (e.g. FLAG, HA) onto your DNA sequence and use that instead of the GFP

its not GFP its GST maybe it doesnt matter. Do you have source read more about it???

[bob1]

Sorry no sources, and you are correct, it doesn't matter whether it is GST or GFP, apart from the use you are going to put it to.  You could try Molecular Cloning a Laboratory manual by Sambrook et al for further info.