GFP标记的短小芽胞杆菌(Bacillus pumilus)转座突变株的构建初探  

Construction of GFP-Labeled Tn5 Insertion Mutants of Bacillus pumilus and gfp Expression Analysis

作　　者：沈新迁[1] 胡晓璐[1] 刘通[1] 孙文良[1,2] 陈云鹏[1] 

机构地区：[1]上海交通大学农业与生物学院,上海200240 [2]中国科学院天津工业生物技术研究所,天津300308

出　　处：《上海交通大学学报（农业科学版）》2012年第4期15-20,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家自然科学基金资助课题(31071721)

摘　　要：以水稻叶面附生菌短小芽孢杆菌(B.pumilus)为研究对象,通过构建pMOD-egfp-Tetr转座复合体,采用电击转化法对短小芽孢杆菌DX01菌株进行了Tn5转座插入诱变,获得了大量的转化子。对转化子进行了遗传稳定性分析和分子鉴定,并且在荧光显微镜下观察到了强烈的绿色荧光,研究结果表明外源DNA已经成功整合进细菌的基因组中,gfp基因在短小芽孢杆菌中实现了组成型表达。Bacillus pumilus is one of the most important biocontrol bacteria for plant.In this study,a pMOD-egfp-Tetr transposition complexes was prepared using Tn5 transposase,and then subjected to the construction of Tn5 transposon insertion mutagenesis for B.pumilus DX01 by electrotransformation.Over one thousand of GFP-tagged transformants were obtained.The analyses of genetic stability test and molecular identification showed that exogenous DNA was successfully integrated into the genome of B.pumilus and strong green fluorescence was observed in transformed cells,which indicated that gfp gene was expressed constitutively in mutants.

关 键 词：短小芽胞杆菌 Tn5转座 电击转化 绿色荧光蛋白 

分 类 号：Q754[生物学—分子生物学]