Re: Mutating CMV promoter

Posted by Thermus aquaticus on Jan 24, 2002 at 19:45
(128.250.250.154)

Re: Mutating CMV promoter (Miryan)

Speaking from experience, site directed mutagenesis is not a trivial exercise. Why do you want to remove the Nde site? If you give me more detail perhaps I can suggest an alternative. Even if the mutation is silent, altering the sequence of a promoter might affect its functioning (depending on where it is) because the binding of RNA polymerase complexes to the promoter is sequence specific.

If it’s a case of ligating something into the vector downstream of the promoter, then using linkers with a different restriction site is a far easier alternative.


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