Hello, people!
I have a classical yeast genetics question that I am not sure.
The problem is I have a heterozygote knock-out diploid cells in h90 background (homothallic strain). {It's a long story why I didn't start with a heterothallic strain.}
Anyway, this mating type switching causes some potential problem. So, I thought about crossing with heterothalic strain to move the knocked-out allele to the heterothallic background. Then, I got confused.
h90 cells are h90 leu1-32,ura4D18, ade6-M210/h90 leu1-32,ura4D18,ade6-M210, KanMX6(G418 selection)::my gene. I want to have cells in the genotype of h90(+) leu1-32, ura4D18, ade6M210/h- leu1-32,ura4D18, ade6M216 KanMX6::my gene. So, I can select the diploids using Ade marker. Then, I would like to isolate haploid cells of h- leu1-32, ura4D18,ade6M216, KanMX6::my gene. My gene is located on 2nd chromosome which mat1,2,3 genes are located. I got more confused at this point whether this can be done or not.
I am looking forward some generous explanations about this.
Sincerely,
Steve K. Cho
Hamon Center for Therapeutic Oncology Research
The University of Texas Southwestern Medical Center at Dallas
NB8.104
5323 Harry Hines Blvd.
Dallas, Tx 75390-8593
Lab: (214)648-4912
Fax: (214)648-4940
e-mail: cho2 at simmons.swmed.edu
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