Hello once again,
Thanks to all those who sent replies to my inquiries regarding integration
of transgenic arrays. I screened 300 plates of cloned F2's and recovered a
strain that gives 100% transmission. The original array consisted of pRF4
and an unc-119::GFP fusion plasmid. All of the animals of this strain
fluoresce and all are right-handed rollers.
I crossed a few of these putative integrants with N2 males, and almost all
the progeny failed to roll. Those that did (only a few) rolled very
weakly. All of the F1's did fluoresce, however, although apparently
slightly weaker than the parent strain. In the F2, the strong right-hand
rol phenotype resegregated, and of those, all fluoresced, and brighter than
the F1's.
It looks like the rol-6D (su1006) has become recessive, maybe due to some
kind of position effect. Either that, or I've integrated the array
_without pRF4_ into a rol gene. (Of course, the apparent recessive nature
of the rol-6 marker makes crossing the 'integrant' easier since the F1
males mate well. Also it will be easier to map it.)
Has anyone else had this happen? As far as I know the F1s should all roll
if it is a true integrant. I'm in the process of backcrossing the F1 males
to the original 'integrant' strain to test for recessive rol.
I'd appreciate any comments.
MM
--
Morris Maduro
Dept of Biological Sciences
University of Alberta
Edmonton, AB Canada Morris_maduro at biology.ualberta.ca
