缬草放线菌中卤代酶类抗生素的筛选及遗传转化系统的建立  

Screening of Halogenated Antibiotics from Actinomyces Valeriana and Establishment of Genetic Transformation System

作　　者：吴佳 贾良辉 WU Jia;JIA Lianghui(Changzhi Medical College)

机构地区：[1]长治医学院医学检验系,046000 [2]西北农林科技大学

出　　处：《长治医学院学报》2021年第4期241-244,共4页Journal of Changzhi Medical College

基　　金：陕西省自然科学基础研究计划-面上项目(2021JM-087)。

摘　　要：目的:从基因水平上挖掘新的卤代酶类抗生素。方法:利用简并引物快速筛选缬草放线菌中含卤代酶类抗生素的目标菌株,并对其进行鉴定。选择萘啶酮酸、氯霉素、潮霉素、卡那霉素、红霉素、庆大霉素、阿普霉素等7种抗生素进行抗生素敏感试验,选择目标菌株的最适抗生素及其最适浓度,构建遗传转化系统。结果:从145株缬草放线菌中筛选到1株含卤代酶类抗生素生物合成基因的菌株AJ56,经鉴定其为链霉菌,在最适抗生素萘啶酮酸和阿普霉素的培养基中构建遗传转化系统,经多次传代,成功构建了菌株AJ56与大肠杆菌S17-1/pSET152遗传转化系统。结论:菌株AJ56遗传转化系统的成功构建为进一步研究其次级代谢产物提供了基础。Objective:New halogenase antibiotics were excavated at the gene level.Methods:Degenerate primers were used to quickly screen and identify the target strains of halogenase antibiotics in valerian actinomycetes.Seven antibiotics including chloramphenicol,hygromycin,kanamycin,erythromycin,gentamicin,apramycin and naphthyridone acid were selected for antibiotic sensitivity test.The optimal antibiotics and their optimal concentrations of the target strains were selected to construct the genetic transformation system.Results:A strain AJ56 containing halogenase antibiotic biosynthesis gene was screened from 145 strains of valerian actinomycetes.It was identified as Streptomyces.The genetic transformation system was constructed in the culture medium of the most suitable antibiotics naphthyridonic acid and Apramycin.After multiple passages,the genetic transformation system between strain AJ56 and E.coli S17-1/pSET152 was successfully constructed.Conclusion:The successful establishment of the genetic transformation system of strain AJ56 provides a basis for further study of its secondary metabolites.

关 键 词：简并引物 放线菌 卤代酶 遗传转化系统 

分 类 号：Q948[生物学—植物学]