Construction of strains for rapid homokaryon purification after integration of constructs at the histidine-3 ( his-3) locus of Neurospora crassa.
We report the construction of histidine-3 (his-3) strains of Neurospora crassa containing the hygromycin B phosphotransferase gene of Escherichia coli (hph(+)) fused in-frame to the herpes simplex virus thymidine kinase gene (tk(+); Lupton et al. 1991), integrated at the his-3 locus. We also report the construction of two ampicillin-resistant and two kanamycin-resistant his-3 gene-replacement vector plasmids. The combined use of these strains and plasmids for his-3-targeted gene integration allows for the rapid identification of homokaryotic transformants containing the expected gene replacement event.
Lee
Dong W
DW
Department of Biology, College of Science, Texas A&M University, Room 415, Building BSBW, College Station, TX 77843-3258, USA.
Haag
Jeremy R
JR
Aramayo
Rodolfo
R
D018014:Gene Transfer Techniques; D006639:Histidine / Q000378:metabolism*; D009492:Neurospora crassa / Q000235:genetics* / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics* / Q000378:metabolism
D006639:Histidine; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase