Lee Dong W, Haag Jeremy R, Aramayo Rodolfo
Department of Biology, College of Science, Texas A&M University, Room 415, Building BSBW, College Station, TX 77843-3258, USA.
Curr Genet. 2003 Apr;43(1):17-23. doi: 10.1007/s00294-002-0366-z. Epub 2003 Feb 18.
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.
我们报告了粗糙脉孢菌组氨酸-3（his-3）菌株的构建，该菌株含有与单纯疱疹病毒胸苷激酶基因（tk(+)；Lupton等人，1991）读框融合的大肠杆菌潮霉素B磷酸转移酶基因（hph(+)），并整合在his-3位点。我们还报告了两种氨苄青霉素抗性和两种卡那霉素抗性的his-3基因置换载体质粒的构建。这些菌株和质粒联合用于靶向his-3的基因整合，能够快速鉴定出含有预期基因置换事件的同核体转化体。
