Varga J, Croft J H
Department of Microbiology, Attila József University, Szeged, Hungary.
Curr Genet. 1994 Apr;25(4):311-7. doi: 10.1007/BF00351483.
Chromosome-substituted haploid segregants were selected from among the benomyl-induced progeny of an interspecific hybrid produced by polyethylene-glycol-induced fusion of protoplasts of an Aspergillus nidulans 'master strain' and an A. quadrilineatus auxotrophic mutant. These segregants were examined by RFLP, RAPD, and isoenzyme analysis. The A. nidulans ribosomal repeat unit was assigned to chromosome V, while the benA and the pyrG genes were assigned to linkage groups VIII and I, respectively, of A. nidulans. None of the other cloned genes tested (gdhA, amdS and 25s rRNA) showed polymorphism between the two parents. The method was also used to assign RAPD markers and isoenzyme bands of beta-arylesterase, phosphatases, NAD-dependent malate dehydrogenase, and cellulase, to A. nidulans chromosomes and/or to their A. quadrilineatus equivalents. The isoenzyme and DNA sequences assigned to chromosomes could be used to saturate the genetic map of A. nidulans, or could serve as starting points for the construction of a genetic map of A. quadrilineatus. No method affording the same possibilities has been described so far in Aspergilli. This chromosome-assay method may be a useful alternative to pulsed-field-gel electrophoretic procedures for the assignment of molecular markers to chromosomes.
从由聚乙二醇诱导的构巢曲霉“主菌株”原生质体与四线曲霉营养缺陷型突变体原生质体融合产生的种间杂种的苯菌灵诱导后代中筛选出染色体替代单倍体分离株。通过限制性片段长度多态性(RFLP)、随机扩增多态性DNA(RAPD)和同工酶分析对这些分离株进行检测。构巢曲霉核糖体重复单位定位于第五条染色体,而benA基因和pyrG基因分别定位于构巢曲霉的第八和第一连锁群。所检测的其他克隆基因(gdhA、amdS和25s rRNA)在两个亲本之间均未显示多态性。该方法还用于将RAPD标记以及β-芳基酯酶、磷酸酶、NAD依赖性苹果酸脱氢酶和纤维素酶的同工酶条带定位于构巢曲霉染色体和/或其四线曲霉对应物上。定位于染色体的同工酶和DNA序列可用于使构巢曲霉的遗传图谱饱和,或可作为构建四线曲霉遗传图谱的起点。目前在曲霉中尚未描述具有相同可能性的方法。这种染色体检测方法可能是一种有用的替代脉冲场凝胶电泳程序的方法,用于将分子标记定位于染色体。