生防链霉菌Men-myco-93-63 nsdB_(mgh)基因的克隆及序列分析  

Cloning and sequence analysis of nsdB_(mgh) gene of biocontrol Streptomyces Men-myco-93-63

作　　者：冀红柳[1] 胡亚亚[1] 沈凤英[2] 李亚宁[1] 刘大群[1] 

机构地区：[1]河北农业大学植物保护学院,河北省植物病虫害生物防治工程技术研究中心,国家北方山区农业工程技术研究中心,河北保定071001 [2]河北北方学院,河北张家口075000

出　　处：《河北农业大学学报》2011年第5期53-58,共6页Journal of Hebei Agricultural University

基　　金：国家自然科学基金项目(31171894);国家高技术研究与发展计划(863)(2011AA10A205);河北省自然科学基金项目(C2011204114)

摘　　要：为了获得生防链霉菌Men-myco-93-63中nsdB基因,根据天蓝色链霉菌A3(2)负调控基因nsdB序列设计引物,扩增Men-myco-93-63基因组DNA,获得1 121 bp的片段,经NPA-PCR对其两端进行延伸,拼接后得到2 073 bp的全长序列,命名为nsdBmgh。经Southern杂交验证,该基因在玫瑰黄链霉菌Men-myco-93-63基因组中确实存在且为单拷贝。该基因在Men-myco-93-63中的发现与克隆,为生防菌的遗传改造和菌株代谢调控途径的研究提供了重要依据。In order to obtain the gene of biocontro Streptomyces Men-myco-93-63,the 1 121 bp gene fragment sequences of genome DNA of Men-myco-93-63 were amplified by the primer sequences designed by S.coelicolor A3（2）.And then,nonspecific primer anchored PCR（NPA-PCR） was used for extending this fragment.After assembling the walking sequence and the original sequence,the full-length sequence was obtained with 2 073 bp,which was named as gene nsdBmgh.The nsdBmgh gene was further confirmed by Southern blot test,and the result showed that there was one copy of nsdBmgh gene in Men-myco-93-63.The detection and cloning of nsdBmgh gene will provide an important basis for further research of genetic transformation and metabolic pathway of biocontrol Streptomyces Men-myco-93-63.

关 键 词：生防链霉菌 nsdB 基因克隆 NPA-PCR 序列分析 

分 类 号：S432.1[农业科学—植物病理学]