Re: Mutating CMV promoter
Posted by Thermus aquaticus on Jan 24, 2002 at 19:45
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 itís 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.