Two consecutive ATG - (Jun/28/2015 )

Hi everyone, 

I requested the following prime (to expression in the pET-29 system):

GAT ATA CAT ATG ATG GCG AAC CAC GCG AAC AAC

(NdeI restriction site highlighted in bold and ATG of the template DNA highlighted in green).

I think the correct would be to remove the initial ATG. Will I have problems using this prime containing two consecutive ATG?

It's always best to initially go with the most native sequence you can.

But more often than not, something small like this will not make a noticeable difference.

But that is why protein chemistry is fun...every protein construct has it's own unique personality, and nobody can predict for you whether a new construct will express, be stable, functional, etc. So only one way to find out.

In the mean time, order the correct primer just in case it does not express as expected.

Hi labtastic, thank you for your help... I will try to express the protein using additional ATG as I requested, I hope there is no problem with the additional methionine at the N-terminal portion. Thank you again