mating possibilities of transgenic Chlamy lines

[Zoltan Kerenyi]

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,

Zoltan

--
Zoltan Kerenyi PhD
Agricultural Biotechnology Center
Szent-Gyorgyi A. u. 4.
H-2100 Godollo, HUNGARY
phone: +36 28 526 100 <%2B36%2020%204555%20349>
email: keren...@gmail.com

[Elizabeth Harris]

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.

Elizabeth Harris