Re: [DIYbio] Expression Problem - fusion protein

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There can be *no* guarantees in biology, of anything.

However, adding two amino acids to the N-terminus of your protein is
unlikely to destroy everything.

Do bear in mind; if the methionine will be excised from the protein,
which itself depends heavily on the second amino, then the second amino
will be the N-terminus of your protein; and the N-terminal amino has a
strong-ish impact on the long-term stability of the protein, with some
aminos (like Valine) being highly stable and others being highly
unstable.

I can't recall offhand but I think likelihood-of-M-excision is higher
if the first few aminos are..simple unbranched/uncharged? Someone else
wanna back me up or correct me here? So Glycine might encourage
methionine processing, and Glycine is probably unstable-ish.

As usual then, the answer is "probably not a big deal but there's all
this random stuff that might go wrong and in the end it might explode
or work amazingly".

On Mon, 7 Oct 2013 07:47:03 -0700 (PDT)
"Mega [Andreas Stuermer]" <masterstorm123@gmail.com> wrote:

> Hi everyone!
>
> For expression of a protein I would need to make it fit to the RBS of
> the vector.
>
> Is it ok, if I attach ATG-GGC (Methionine and Glycine) to the
> N-Terminus?
>
> So the fusion protein would be Met-Gly-Met-rest of Protein
>
> Is there a guarantee that the protein still works then?
>
> Best,
> Andreas
>