[Fwd: Binary vector dependent unintended misexpression]

Nobody nobody at hgmp.mrc.ac.uk
Wed Nov 21 04:57:07 EST 2001


Hi,

I don't remember my pCambia's, but my guess would be read through from some
other promoter upstream, possibly 35S, as I think they all have that.
Terminators are not really terminators, but just 'braking zones' for the RNA
polymerase to fall off, and a lot of the artificial ones are very short.  It
is noteworthy that pBIN19 effectively has a very long terminator after the
nos::kan gene, due to its numerous erroneous insertions - which happen to
enhance function.

If the added gene had a very long promoter fragment, it might help the RNA
polymerase from the previous gene fall off (is it very short?), but a better
be to would be to add an extra spacer element of some sort.  Not less that 5=
00
bp would be my guess.  Or just use another vector...!


Cheers,

Trevor Fenning.

Jim Tokuhisa wrote:

>=20 interesting!  Any ideas what might be happening.
>
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