从兼性甲基营养菌假单胞菌属AM1菌株中克隆苹果酰辅酶A裂合酶基因的分子克隆。
Molecular cloning of a malyl coenzyme A lyase gene from Pseudomonas sp. strain AM1, a facultative methylotroph.
作者信息
Fulton G L, Nunn D N, Lidstrom M E
出版信息
J Bacteriol. 1984 Nov;160(2):718-23. doi: 10.1128/jb.160.2.718-723.1984.
Abstract
A genomic library containing HindIII partial digests of Pseudomonas sp. strain AM1 DNA was constructed in the broad-host-range cosmid pVK100. PCT57, a Pseudomonas sp. strain AM1 methanol mutant deficient in malyl coenzyme A lyase activity, was complemented to a methanol-positive phenotype by mobilization of the pVK100 library into PCT57 recipients with the ColE1/RK2 mobilizing plasmid pRK2013. Six different complemented isolates all contained a recombinant plasmid carrying the same 19.6-kilobase-pair Pseudomonas sp. strain AM1 DNA insert. Subcloning and complementation analysis demonstrated that the gene deficient in PCT57 (mcl-1) was located in a 1.6-kilobase-pair region within a 7.4-kilobase-pair EcoRI-HindIII fragment.
摘要
用广宿主范围的黏粒pVK100构建了一个包含假单胞菌属菌株AM1 DNA的HindIII部分酶切片段的基因组文库。PCT57是一株缺乏苹果酰辅酶A裂合酶活性的假单胞菌属菌株AM1甲醇突变体，通过用ColE1/RK2转移质粒pRK2013将pVK100文库转移到PCT57受体菌中，使其恢复为甲醇阳性表型。六个不同的互补分离株均含有一个携带相同19.6千碱基对假单胞菌属菌株AM1 DNA插入片段的重组质粒。亚克隆和互补分析表明，PCT57中缺陷的基因（mcl-1）位于一个7.4千碱基对的EcoRI-HindIII片段内的一个1.6千碱基对区域。
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