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参与转运核糖核酸（tRNA）中δ2-异戊烯基腺苷形成的大肠杆菌miaA基因的分子克隆

Molecular cloning of the Escherichia coli miaA gene involved in the formation of delta 2-isopentenyl adenosine in tRNA.

作者信息

Caillet J, Droogmans L

机构信息

Institut de Biologie Physico-Chimique, Paris, France.

出版信息

J Bacteriol. 1988 Sep;170(9):4147-52. doi: 10.1128/jb.170.9.4147-4152.1988.

DOI:10.1128/jb.170.9.4147-4152.1988
PMID:3045085
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC211421/
Abstract

Escherichia coli mia strains were shown to lack delta 2-isopentenylpyrophosphate transferase activity, the first step in the synthesis of the 2-methylthio derivative of 6-(delta 2-isopentenyl) adenosine (ms2i6A). A double mutant, rpsL (Smp) miaA, was streptomycin dependent. The wild-type miaA gene was cloned by selecting for lambda recombinant bacteriophage which eliminated the streptomycin-dependent phenotype and was subsequently recloned into plasmid vectors. The cloned miaA gene restored the ms2i6A modification to tRNA. The miaA gene mapped to 95 min on the E. coli map, and we propose the order mutL-miaA-hflA-purA.

摘要

已证明大肠杆菌mia菌株缺乏δ2-异戊烯基焦磷酸转移酶活性，这是6-(δ2-异戊烯基)腺苷(ms2i6A)的2-甲硫基衍生物合成的第一步。双突变体rpsL(Smp)miaA对链霉素有依赖性。通过选择消除链霉素依赖性表型的λ重组噬菌体克隆了野生型miaA基因，随后将其重新克隆到质粒载体中。克隆的miaA基因恢复了tRNA的ms2i6A修饰。miaA基因在大肠杆菌染色体图谱上定位于95分钟处，我们提出mutL-miaA-hflA-purA的顺序。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27c5/211421/53534af6c53e/jbacter00187-0370-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27c5/211421/53534af6c53e/jbacter00187-0370-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/27c5/211421/53534af6c53e/jbacter00187-0370-a.jpg

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