Uitzetter J H, Bos C J, Visser J
J Gen Microbiol. 1986 May;132(5):1167-72. doi: 10.1099/00221287-132-5-1167.
A selective method for the isolation of Aspergillus nidulans mutants defective in the pyruvate dehydrogenase complex was devised. The essential steps in the procedure were a mutagenic treatment of conidia with X-rays to about 50% survival, followed by filtration enrichment in minimal medium with D-galacturonate as sole carbon source, and rescue on complete medium with acetate. The mutants thus isolated were phenotypically characterized on the basis of growth tests, and different genotypes were assigned on the basis of complementation tests. The majority of the mutants that were unable to utilize galacturonate were defective in one of the components of the pyruvate dehydrogenase complex. In addition, mutants defective in pyruvate carboxylase, mutants defective in glycerol catabolism and some novel mutants which were only unable to use D-galacturonate as carbon source were found. At least two genes were shown to be involved in D-galacturonate metabolism.
设计了一种用于分离丙酮酸脱氢酶复合体缺陷型构巢曲霉突变体的筛选方法。该方法的关键步骤包括:用X射线对分生孢子进行诱变处理,使其存活率约为50%,随后在以D - 半乳糖醛酸作为唯一碳源的基本培养基中进行过滤富集,然后在添加乙酸盐的完全培养基上进行挽救。如此分离得到的突变体通过生长测试进行表型特征分析,并通过互补测试确定不同的基因型。大多数无法利用半乳糖醛酸的突变体在丙酮酸脱氢酶复合体的某个组分中存在缺陷。此外,还发现了丙酮酸羧化酶缺陷型突变体、甘油分解代谢缺陷型突变体以及一些仅无法将D - 半乳糖醛酸用作碳源的新型突变体。至少有两个基因参与了D - 半乳糖醛酸的代谢。