Re: [DIYbio] PolyA tails

I'm not seeing any long stretch of polyA or polyT anywhere in that
plasmid. In snapgene the feature is called "CaMV poly(A) signal" but
it doesn't actually have a stretch of A's or T's, aside from the 4
consecutive A's starting at position 2961. But of course that would
be transcribed to UUUU.

-cory

On Thu, Mar 20, 2014 at 12:29 PM, Sebastian Cocioba <scocioba@gmail.com> wrote:
> Yes, but also I noticed that the CaMV in pGreenII 0049 is terminated
> with a polyA instead of the usual tnos or other defined terminator. Is
> that a kind of preadenylation or just coincidental that it also
> terminates CaMV well. SnapGene has the plasmid in its vector directory
> and it anotated a polyCap of sorts in the DNA.
>
> Sebastian S. Cocioba
> CEO & Founder
> New York Botanics, LLC
> Plant Biotech R&D From: Cory Tobin
> Sent: 3/20/2014 3:24 PM
> To: diybio
> Subject: Re: [DIYbio] PolyA tails
> Maybe but maybe not. The poly(a) tail does more than just prevent
> degradation. It interacts with the ribosome and the 5' UTR complex to
> control the efficiency/rate of translation. Making the tail too long
> may disrupt translation, although I can't find any evidence that it
> does in the literature. I'm just speculating that there may be
> unintended consequences to lengthening the tail.
> Here's a good review on the subject:
> http://www.ncbi.nlm.nih.gov/pubmed/22664985 email me if you need the
> pdf.
>
> Also, I don't know of any way to actually increase the length of the
> tail since it's not coded in the DNA. Well, it sort of is. The
> affinity of the poly(A) polymerase, CPSF and PABII for the mRNA
> probably effect the length of the tail and those proteins are coded
> for in the DNA. But you know what I mean. There isn't a stretch of
> poly(T) in the DNA downstream of the CDS that get transcribed into the
> tail.
>
> I think there's ways of extending it in vitro. So if you're planning
> on injecting mRNA into a cell, that could possibly work. But I
> figured you were talking about engineering a (plant?) cell to produce
> a long-living mRNA.
>
> -cory
>
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--
Cory Tobin
Graduate Student
Caltech :: Biology
ctobin@caltech.edu
805-443-4702
1200 E. California Blvd MC156-29
Pasadena CA, 91125

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