Most efficient PCR strategy for generating fusion (chimeric) proteins ?

ab via methods%40net.bio.net (by anirbn from gmail.com)
Thu Oct 9 22:54:33 EST 2008


Dear all,

I have to generate several chimeric proteins where I will have to
insert about 50 aa residues of one protein in the middle of another. I
was searching through the archives and found the Emerman, Deminie
protocol ( J.Virol , July 1993 p.6499-6506). Basically one has to do
four PCR reactions followed by a restriction digest and ligation. Is
this still the most efficient way to generate chimeric proteins where
the insertion size is relatively big ? Any help will be really
appreciated.

Best regards,

AB.



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