Alic M, Akileswaran L, Gold M H
Department of Chemistry, Biochemistry, and Molecular Biology, Oregon Graduate Institute of Science and Technology, Beaverton 97006.
Gene. 1993 Dec 22;136(1-2):307-11. doi: 10.1016/0378-1119(93)90485-l.
The ability to carry out gene replacements and gene targeting in the lignin-degrading basidiomycete fungus, Phanerochaete chrysosporium, would facilitate studies on the roles and regulation of various components of its lignindegrading system. A plasmid consisting of the P. chrysosporium ura3 gene (encoding orotidylate decarboxylase) interrupted with the Schizophyllum commune ade2 gene (encoding an adenine biosynthetic enzyme) was used to transform the P. chrysosporium ade2 strain to adenine prototrophy with selection on 5-fluoroorotic acid for inactivation of the ura3 gene. Stable Ade+Ura- strains were obtained at a frequency of approximately one transformant per microgram of DNA. In all of the Ade+Ura- transformants examined by Southern analysis, the chromosomal ura3 locus had been replaced by the plasmid insert.
在木质素降解担子菌黄孢原毛平革菌中进行基因替换和基因靶向的能力，将有助于研究其木质素降解系统各组分的作用及调控。一个由被裂褶菌ade2基因（编码腺嘌呤生物合成酶）中断的黄孢原毛平革菌ura3基因（编码乳清酸脱羧酶）组成的质粒，被用于将黄孢原毛平革菌ade2菌株转化为腺嘌呤原养型，并在5-氟乳清酸上进行筛选以使ura3基因失活。以每微克DNA约一个转化体的频率获得了稳定的Ade⁺Ura⁻菌株。在通过Southern分析检测的所有Ade⁺Ura⁻转化体中，染色体ura3位点已被质粒插入片段所取代。
