猪霍乱沙门氏菌C500株crp^-/gfp^＋缺失株的构建及鉴定  

Construction and Characterization of crp^-/gfp^+ Mutant of Salmonella choleraesuis C500 Strain

作　　者：徐引弟[1] 郭爱珍[1] 陈焕春[1] 

机构地区：[1]华中农业大学农业微生物学国家重点实验室

出　　处：《农业生物技术学报》2008年第2期196-201,共6页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金资助项目(No.30571386);国家高技术研究与发展计划(863)项目(No.2006AA10A206)资助

摘　　要：构建了猪霍乱沙门氏菌(Salmonella choleraesuis)C500株的crp缺失与插入gfp基因的crp-/gfp+缺失株。首先构建含缺失320bp的crp基因并插入gfp基因的重组自杀性质粒,与C500接合转移,两步法筛选无抗性的crp-/gfp+缺失株,经PCR、测序鉴定和荧光显微镜观察证实在C500的基因组上成功缺失了crp基因并正确插入了gfp基因。Salmonella choleraesuis C500 strain crp^-/gfp^＋ mutant obtained by deleting crp gene and inserting gfp gene was constructed. Firstly, the recombination suicide vector with 320 bp-deleted crp gene and 720 bp-insert gfp gene was constructed and conjugated with C500. The unmarked crp deleted and gfp inserted strain was selected by a two-step method. The crp deletion and gfp insertion on the genome were determined by PCR, sequencing and fluorescence observation with microscope, indicating that the crp^-/gfp^＋ mutant was successfully constructed.

关 键 词：猪霍乱沙门氏菌C500株 crp基因 GFP基因 基因缺失 重组自杀性质粒 

分 类 号：S188[农业科学—农业基础科学]