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Smallest mRNA length that can be translated


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#1 Ahrenhase

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Posted 04 November 2013 - 09:59 AM

Trying to generate minigenes for in vitro transcription and translation.  



#2 bob1

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Posted 06 November 2013 - 12:55 PM

Including UTRs?  I would guess (and it is a real guess) that the minimum will be defined by a ribosome binding site, the start codon and at least 1 amino acid (I suspect you would need more than this) before the stop codon.  How you would tell if there has been translation would be quite difficult, though you could perhaps do it through size exclusion chromatography or MS.



#3 Ahrenhase

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Posted 25 November 2013 - 08:51 PM

Including UTRs?  I would guess (and it is a real guess) that the minimum will be defined by a ribosome binding site, the start codon and at least 1 amino acid (I suspect you would need more than this) before the stop codon.  How you would tell if there has been translation would be quite difficult, though you could perhaps do it through size exclusion chromatography or MS.

From promega, they suggest a 5' UTR of 15-100 bp for maximal translation.  I will have this + ~100 amino acids encoding aftwards, which I'm confident will translate. I will report back if this works. 



#4 bob1

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Posted 25 November 2013 - 11:14 PM

That should work fine - after all you can get short tags (3x Flag for example) to express off plasmids.






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