Yaffe M P, Jensen R E, Guido E C
Department of Biology, University of California, San Diego, La Jolla 92093.
J Biol Chem. 1989 Dec 15;264(35):21091-6.
As part of an analysis of the function and assembly of the mitochondrial outer membrane, we have cloned and characterized the yeast gene encoding a 45-kDa polypeptide (OM45) which is a major constituent of this membrane. The nuclear gene was isolated by immunological screening of plaques of recombinant phage lambda gt11 containing fragments of yeast genomic DNA using an antibody against OM45. Determination of the nucleotide sequence of the DNA fragment isolated by this approach revealed a single open reading frame of 1179 base pairs which encodes a protein having a predicted molecular mass of 44.6-kDa. Disruption of the OM45 gene in haploid yeast cells eliminated the expression of OM45. The mutant strain showed no apparent defect in cell viability, growth, mitochondrial function, or mitochondrial protein import.
作为对线粒体外膜功能和组装分析的一部分，我们克隆并鉴定了编码一种45 kDa多肽（OM45）的酵母基因，该多肽是该膜的主要成分。通过使用抗OM45抗体对含有酵母基因组DNA片段的重组噬菌体λgt11的噬菌斑进行免疫筛选，分离出了该核基因。通过这种方法分离的DNA片段的核苷酸序列测定揭示了一个1179个碱基对的单一开放阅读框，其编码一种预测分子量为44.6 kDa的蛋白质。单倍体酵母细胞中OM45基因的破坏消除了OM45的表达。突变菌株在细胞活力、生长、线粒体功能或线粒体蛋白导入方面没有明显缺陷。
