Edwards D L, Belsole D M, Guzik H J, Unger B W
J Bacteriol. 1979 Feb;137(2):900-4. doi: 10.1128/jb.137.2.900-904.1979.
A method is described which permits the selection of mutants of Neurospora crassa that are deficient in succinic dehydrogenase activity. The method relies on the observation that succinic dehydrogenase-deficient strains fail to reduce the dye nitrotetrazolium blue when overlaid with the dye in the presence of succinate and phenazine methosulfate. Wild-type colonies reduced the dye and turned blue, whereas mutant colonies remained colorless. In this communication we present studies of a mutant, SDH-1, isolated by this method. The mutant had 18% of the succinic dehydrogenase activity of the parent strain used in the mutation experiments as determined from the ratio of Vmax activities obtained from Lineweaver-Burk plots. The SDH-1 mutant segregated in a Mendelian manner when back-crossed to its parent strain. Succinate oxidase activity in SDH-1 was low and was markedly inhibited by adenosine 5'-diphosphate. The succinate oxidase activity of the parent strain was high and was not affected by the presence of adenosine 5'-diphosphate.
本文描述了一种方法,该方法可用于筛选出琥珀酸脱氢酶活性缺陷的粗糙脉孢菌突变体。该方法基于这样的观察结果:当在琥珀酸盐和吩嗪硫酸甲酯存在的情况下,将缺乏琥珀酸脱氢酶的菌株与染料硝基四氮唑蓝覆盖时,这些菌株无法还原该染料。野生型菌落会还原染料并变成蓝色,而突变型菌落则保持无色。在本通讯中,我们展示了通过该方法分离出的一个突变体SDH-1的研究。根据从Lineweaver-Burk图获得的Vmax活性比率测定,该突变体的琥珀酸脱氢酶活性为用于突变实验的亲本菌株的18%。当与亲本菌株回交时,SDH-1突变体以孟德尔方式分离。SDH-1中的琥珀酸氧化酶活性较低,并且被5'-二磷酸腺苷显著抑制。亲本菌株的琥珀酸氧化酶活性较高,并且不受5'-二磷酸腺苷存在的影响。