Ferguson D O, Rice M C, Rendi M H, Kotani H, Kmiec E B, Holloman W K
Hearst Microbiology Research Center, Department of Microbiology, Cornell University Medical College, New York, New York 10021, USA.
Genetics. 1997 Feb;145(2):243-51. doi: 10.1093/genetics/145.2.243.
A gene encoding a Ustilago maydis Rad51 orthologue has been isolated, rad51-1, a mutant constructed by disrupting the gene, was as sensitive to killing by ultraviolet light and gamma radiation as the rec2-1 mutant and slightly more sensitive to killing by methyl methanesulfonate. There was no suppression of killing by ultraviolet light when a rec2-1 strain was transformed with a multicopy plasmid containing RAD51, nor was there suppression when rad51-1 was transformed with a multicopy plasmid containing REC2. Recombination proficiency as measured by a gap repair assay was diminished in both rec2-1 and rad51-1 strains. In rec2-1 the frequency of recombination was decreased, but the spectrum of events was similar to that observed in wild type, while in rad51-1 the frequency as well as the spectrum of recombination events were different. Studies with the rec2-1 rad51-1 double mutant indicated that there was epistasis in the action of REC2 and RAD51 in certain repair and recombination functions, but some measure of independent action in other functions.
已分离出一个编码玉米黑粉菌Rad51直系同源物的基因,通过破坏该基因构建的突变体rad51-1,对紫外线和γ辐射杀伤的敏感性与rec2-1突变体相同,对甲基磺酸甲酯杀伤的敏感性略高。当用含有RAD51的多拷贝质粒转化rec2-1菌株时,紫外线杀伤没有得到抑制,当用含有REC2的多拷贝质粒转化rad51-1时也没有抑制作用。通过缺口修复试验测定的重组能力在rec2-1和rad51-1菌株中均降低。在rec2-1中,重组频率降低,但事件谱与野生型中观察到的值相似,而在rad51-1中,重组事件的频率和谱均不同。对rec2-1 rad51-1双突变体的研究表明,REC2和RAD51在某些修复和重组功能中的作用存在上位性,但在其他功能中存在一定程度的独立作用。