[Chlamydomonas] mating possibilities of transgenic Chlamy lines
(by chlamy from duke.edu)
Wed Feb 1 09:09:45 EST 2012
Cultures of wall-deficient mutants typically contain many cells with no flagella, and hence are likely to give poor mating efficiencies. Also, cw products of crosses often do not grow after zygospore germination.
You will probably get better results if you use strains with normal walls, using autolysin to remove the walls prior to transformation.
See http://www.chlamy.org/methods/autolysin2.html for a protocol for preparing this enzyme.
On Feb 1, 2012, at 5:02 AM, Zoltan Kerenyi wrote:
> Dear Members,
> I am planning now a project which targets cytoplasmic mRNA degradation
> pathways mainly the deadenylation step. I plan to make transgenic Chlamy
> lines containing reporter constructs and other lines in which the key genes
> of deadenylation are knocked down. I would like to cross the two types of
> transgenic lines in order to get strains that express the reporter gene and
> the knock down construct at the same time in the same cells.
> I would like to know whether mating of two transgenic lines (cell wall
> deficient mutants) possible is or not.
> Thanks in advance,
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