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For E. coli translation of an mRNA, do you need space between the stop codon and


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#1 Brian Jackson

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Posted 03 November 2011 - 01:52 PM

I am trying to design a forward primer to amplify an ORF to make an mRNA to express a protein without the fusion tags in a bacterial system. So the vector goes something like this

[Promoters][Ribosome binding sequence][fusion tags (V5/His x6)][Stop codon]*[Ribosome binding sequence][9-10 nucleotides][ATG-my protein][Stop codon].

My question is: Do I need to add any spacer nucleotides between the first stop codon and the next ribosome binding sequence (at the *), or can they be right next to each other?

#2 pDNA

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Posted 05 November 2011 - 04:46 AM

can be right after each other ...e.g. see the lac operon!

so no special spacer needed!

Regards,
p

#3 Brian Jackson

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Posted 05 November 2011 - 10:45 AM

Great, thanks!




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