Protocol Online logo
Top : New Forum Archives (2009-): : Protein Expression and Purification

protein expression by changing start codon - (Mar/09/2013 )

Hi all,
I am grying to clone a gene that begins with a GTG strat codon rather than an ATG. Usually i ann an Nde1 restriction site at the 5'end of the gene to be PCR amplified . Hence i remove the ATG start codon from my gene during primer design and insert the Nde1 restriction site at 5'end. In the same way is it possible to remove the GTG start codon and add the Nde1 site that carries the ATG ? will there be any effects on protein expression?
pls help
Thank you


Sure it is possible, in exactly the manner you described - just add the ATG in the place of the GTG in the primer.

There could be problems with expression, but it will probably depend on the protein you are trying to express.