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牛心线粒体ATP合酶的δ亚基。借助聚合酶链反应克隆其导入前体的互补DNA序列。

The delta-subunit of ATP synthase from bovine heart mitochondria. Complementary DNA sequence of its import precursor cloned with the aid of the polymerase chain reaction.

作者信息

Runswick M J, Medd S M, Walker J E

机构信息

M.R.C. Laboratory of Molecular Biology, Cambridge, U.K.

出版信息

Biochem J. 1990 Mar 1;266(2):421-6. doi: 10.1042/bj2660421.

Abstract

The delta-subunit of ATP synthase from bovine heart mitochondria is part of the extrinsic membrane domain, F1-ATPase. The mature protein is 146 amino acids in length and its function is obscure. It is encoded by a nuclear gene and is imported into the organelle. Two mixtures of oligonucleotides 17 bases long, designed on the basis of the known protein sequence, have been synthesized and employed as primers on bovine cDNA in the polymerase chain reaction. By this means a segment of bovine cDNA encoding part of the delta-subunit has been amplified, and this DNA segment has been employed to identify related cDNA clones in a library. These clones encode the mitochondrial import precursor of the delta-subunit; the protein sequence of the mature protein deduced from it is exactly the same as that determined earlier by direct sequence analysis. The clones have also been used to show that both the bovine and human genomes seem to contain a single gene for the delta-subunit.

摘要

牛心线粒体ATP合酶的δ亚基是外在膜结构域F1 - ATP酶的一部分。成熟蛋白长度为146个氨基酸,其功能尚不清楚。它由核基因编码并被导入细胞器。根据已知的蛋白质序列设计了两种长度为17个碱基的寡核苷酸混合物,并在聚合酶链反应中用作牛cDNA的引物。通过这种方法,编码δ亚基部分的一段牛cDNA被扩增出来,并且该DNA片段已被用于在文库中鉴定相关的cDNA克隆。这些克隆编码δ亚基的线粒体导入前体;由此推导的成熟蛋白的蛋白质序列与先前通过直接序列分析确定的序列完全相同。这些克隆还被用于表明牛和人类基因组似乎都只包含一个δ亚基基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/947e/1131148/330513086403/biochemj00188-0114-a.jpg

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