Harkness T A, Rothery R A, Weiner J H, Werner S, Azevedo J E, Videira A, Nargang F E
Department of Biological Sciences, University of Alberta, Edmonton, Canada.
Curr Genet. 1995 Mar;27(4):339-50. doi: 10.1007/BF00352103.
We have used the procedure of sheltered RIP to generate mutants of the 78-kDa protein of the peripheral arm of Neurospora crassa complex I. The nuclei containing the mutations were initially isolated as one component of a heterokaryon but subsequent analysis showed that nuclei containing null alleles of the gene could be propagated as homokaryons. This demonstrates that the gene does not serve an essential function. Sequence analysis of one allele shows that 61 transition mutations were created resulting in 39 amino-acid changes including the introduction of four stop codons. Mutant strains grow at a slower rate than wild-type and exhibit a decrease in the production of conidia. Electron paramagnetic spectroscopy of mutant mitochondria suggest that they are deficient in Fe-S clusters N-1, N-3, and N-4.
我们利用隐蔽性核糖体失活蛋白（RIP）程序生成了粗糙脉孢菌复合体I外周臂78 kDa蛋白的突变体。最初，含有突变的细胞核作为异核体的一个组分被分离出来，但后续分析表明，含有该基因无效等位基因的细胞核可以作为同核体进行繁殖。这表明该基因不具有必需功能。对一个等位基因的序列分析显示，产生了61个转换突变，导致39个氨基酸变化，包括引入了四个终止密码子。突变菌株的生长速度比野生型慢，分生孢子产量降低。突变线粒体的电子顺磁共振光谱表明，它们缺乏铁硫簇N-1、N-3和N-4。
