Molecular cloning of the gene cluster for lariatin biosynthesis of Rhodococcus jostii K01-B0171
Molecular cloning of the gene cluster for lariatin biosynthesis of Rhodococcus jostii K01-B0171
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DOI：
10.1007/s00253-012-3973-8
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发表时间：
2012-07-01
影响因子：
5
通讯作者：
Tomoda, Hiroshi
中科院分区：
文献类型：
--
作者：
Inokoshi, Junji;Matsuhama, Maki;Tomoda, Hiroshi
从革兰氏阳性菌约氏红球菌K01-B0171中克隆了具有独特“套索”结构的抗分枝杆菌肽抗生素lariatins A和B的生物合成基因簇。利用IS1415衍生物进行随机转座诱变，鉴定了一个参与幼虫素生物合成的染色体位点，获得了6个独立的幼虫素不产生变异。任意PCR结果显示，其中一个插入位点位于与幼虫生物合成有关的区域附近。以疟蚊基因为探针，利用集落杂交技术筛选了白蛉K01-B0171的基因组文库，获得了2个克隆。对这些克隆的序列分析显示，lariatin生物合成基因簇长约4.5 kb，由5个开放阅读框（larA ~ larE）组成。提出线性前驱体LarA经LarB、LarC和LarD加工，成熟后的LarA经LarE输出。
The biosynthetic gene cluster for lariatins A and B, anti-mycobacterial peptide antibiotics with a unique "lasso" structure, was cloned from Gram-positive bacterium Rhodococcus jostii K01-B0171. Random transposition mutagenesis using IS1415 derivative was carried out to identify a chromosomal locus involved in lariatin biosynthesis and six independent lariatin non-producing variants were obtained. Arbitrary PCR revealed that one insertion was located near the region involved in lariatin biosynthesis. Using the lariatin gene as a probe, a genomic library of R. jostii K01-B0171 was screened by colony hybridization, and two clones were obtained. Sequence analysis of these clones revealed that the gene cluster for lariatin biosynthesis spanning about 4.5 kb consisted of five open reading frames (larA to larE). We proposed that the linear precursor LarA is processed by LarB, LarC, and LarD, and the mature lariatin is exported by LarE.