产色链霉菌中参与多氧霉素生物合成的基因sanO的克隆与特性分析

Cloning and characterization of sanO, a gene involved in nikkomycin biosynthesis in Streptomyces ansochromogenes.

作者信息

Wang G, Nie L, Tan H

机构信息

Department of Molecular Microbiology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.

出版信息

Lett Appl Microbiol. 2003;37(6):452-7. doi: 10.1046/j.1472-765x.2003.01426.x.

DOI:10.1046/j.1472-765x.2003.01426.x

PMID:14633098

Abstract

AIMS

To clone and characterize sanO, a gene involved in the biosynthesis of nikkomycins in Streptomyces ansochromogenes.

METHODS AND RESULTS

A 4.5-kb BamHI-KpnI fragment was cloned and sequenced. Sequence analysis revealed that this fragment contains three complete open reading frames. The largest one with 2034 bp was designated sanO, which encodes a protein consisting of 667 amino acids with high similarity to module of peptide synthetase. sanO disruption mutants produced no nikkomycin X, but formed nikkomycin Z at almost the same level of the wild-type strain. The production of nikkomycin X can be recovered by genetic complementation of sanO disruption mutants. Primer extension also revealed that transcription start point(tsp) of sanO was localized 87 bp upstream of the potential start codon (GTG).

CONCLUSIONS

sanO was essential for the biosynthesis of nikkomycin X.

SIGNIFICANCE AND IMPACT OF THE STUDY

Nikkomycins have received increased interest of study because of their prospective application in agriculture and medicine. Cloning and characterization of genes involved in the nikkomycin biosynthesis will help to elucidate the whole biosynthetic pathway of nikkomycins.

摘要

目的

克隆并鉴定参与产色链霉菌中多氧霉素生物合成的基因sanO。

方法与结果

克隆并测序了一个4.5 kb的BamHI - KpnI片段。序列分析表明该片段包含三个完整的开放阅读框。其中最大的一个为2034 bp，被命名为sanO，它编码一个由667个氨基酸组成的蛋白质，与肽合成酶模块具有高度相似性。sanO基因敲除突变体不产生多氧霉素X，但产生的多氧霉素Z与野生型菌株几乎处于相同水平。多氧霉素X的产量可通过sanO基因敲除突变体的遗传互补得以恢复。引物延伸分析还表明，sanO的转录起始点（tsp）位于潜在起始密码子（GTG）上游87 bp处。