Do I need to always put ATG as first codon when I make a C- terminal mutant?

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Michael J. Benedik

If you want the protein to be expressed then you do need an appropriate start codon (usually ATG) correctly positioned after a ribosome binding site. Otherwise you won't get any protein expression.

Manuele Martinelli

Dear Qinhong Wang

what do you mean for C-terminal mutant?

A single point mutation at the C-term? The production of a shorter C-terminal fragment which miss the N-term?

best

Manuele