RAPD法对Streptomyces+gilvosporeus+基因组重排前后差异条带的分析研究.pdfVIP

RAPD 法对Streptomyces gilvosporeus 基因组重排前后差异条带的分析 （天津科技大学生物工程学院 天津市工业微生物重点实验室，天津 300457 ） *1 王敏 刘丹 骆健美 刘峰 摘 要：以SDS 法提取褐黄孢链霉菌基因组DNA ，在已获得最佳随机扩增多态性（RAPD ） 条件的基础上，以出发菌株为模板，从20 种随机引物中筛选出了一条能在该链霉菌中扩增 出清晰，稳定片段的引物，即引物S1458。采用该引物对Streptomyces gilvosporeus 基因组重 排后菌株与出发菌株进行比较分析，发现该引物能检测到重排后菌株和出发菌株的差异，即 重排后菌株出现了一条 1500bp 左右的特异性条带。将两菌株间存在的差异条带在大肠杆菌 DH5 α中克隆，发现所得部分序列与GenBank 中几种链霉菌中存在的MarR 家族转录调控 因子存在很高的同源性。 关键词：随机引物扩增多态性；褐黄孢链霉菌；基因组重排；MarR 家族转录调控因子 中图分类号：Q7 文献标识码：A Analyzing DNA difference of shuffling-induced Streptomyces gilvosporeus by RAPD ( Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science Technology, Tianjin 300457, China) WANG Min LIU Dan LUO Jian-Mei LIU Feng Abstract ：The SDS method was used to extract genomic DNA from Streptomyces gilvosporeus. Based on the optimum RAPD patterns, one 10 mer primer, S1458 was screened out from 20 primes by using initial strain as DNA template with good repeatability and polymorphism and then was used to analyze DNA difference of shuffling-induced and initial strains. It was found that one specific fragment that is about 1500bp was obtained in shuffling-induced strain and cloned in E.Coli DH5. Partial sequence of the different gene band had a high homology with the 基金项目：天津市应用基础研究计划面上项目(07JCYBJC07800)， 天津科技大学引进人才科研启动基金， IFS 国际科学基金(F/4239-1) ． *通讯作者：骆健美（1978- ），女，博士，副教授，Tel:022 E-mail:luojianmei@tust.edu.cn MarR-family transcription regulator factor existing in several streptomyces. Key words ：RAPD; Streptomyces gilvosporeus; Genome shuffling; MarR-family transcription regulator factor 随机扩增多态性DNA(random amplified polymorphic