1株来自海洋的芽胞杆菌dhs-330转座突变库的构建及表面活性相关基因的克隆

Construction of Transposon Mutation Library from a Marine Bacillus sp.dhs-330 and The Cloning of Surface Activity Related Genes

为研究产生物表面活性剂的海洋芽胞杆菌dhs-330合成活性产物的分子机制,应用质粒p IC333介导的mini-Tn10转座子随机突变技术,构建了芽胞杆菌dhs-330的突变体库。通过表面活性测定和反向PCR克隆,从300个突变株中筛选出产表面活性剂水平提高的突变株2株,分别在ycs G和yvk C基因发生插入突变;表面活性降低的突变株4株,分别在fen C、yrk F、kin E和sig D基因发生插入突变。这些基因可能与芽胞杆菌dhs-330中表面活性剂的合成代谢和调控有关。

In order to study the molecular mechanism of bio-surfactant synthesis in a marine biosurfactant-producing Bacillus dhs-330 strain,the dhs-330 mutation library was built via mini-Tn10 transposon random mutation technique mediated by plasmid p IC333 applied in this study. Through the measurement of surface activity and inverse PCR cloning,two strains that enhanced their production of surfactant were screened with insertion mutation on yes G and yvk C genes had happened respectively from 300 mutants; and four mutants that decreased their surface activity,with insertion mutation on fen C,yrk F,kin E and sig D genes had happened respectively. It was speculated that these genes may be related to the synthetic metabolism and regulation of biosurfactant in Bacillus sp. dhs-330.