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Cloning a fusion protein, do I remove the ATG codon.

N terminal fusion ATG primer desgin

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#1 Teapot_11



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Posted 15 August 2014 - 10:50 AM

Hi all,

I have a dilemma if anyone was kind and knowledgeable enough.

I am cloning a N-terminal fusion protein, and I am designing my primers so that I have to remove the ATG codon of my gene of interest.


My question --> is it appropriate to remove the ATG from your protein and rely on the transcription of you fused protein within the vector?


I am little stumped on why you have to remove the ATG in the first place?


Would appreciate some ideas.



Yours Sincerely Liam Tegg

#2 bob1


    Thelymitra pulchella

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Posted 15 August 2014 - 02:48 PM

You don't need to, but if you are using a short tag you run the risk that the RNA pol will skip over the tag and start transcription at the internal ATG. To get around this, you can add a Kozak (or Shine-Delgarno, depending on application) sequence at the ATG for the tag.

Also tagged with one or more of these keywords: N terminal fusion, ATG, primer desgin

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