志贺菌福氏2a信号标签诱变突变体文库的构建  

Construction of mutant library of Shigella flexneri2a by signature-tagg ed mu tagenesis

作　　者：姚潇[1] 王恒梁[1] 杨伯伦[2] 史兆兴[1] 冯尔玲[1] 苏国富[1] 黄留玉[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071 [2]西安交通大学环境与化学工程学院,西安710049

出　　处：《生物技术通讯》2003年第1期25-28,共4页Letters in Biotechnology

基　　金：首都二四八重大创新工程(H010210360119);国家重点基础研究发展规划(973项目)(G1999054103)

摘　　要：以合成的单链序列特异性标签为模板,通过PCR得到双链DNA标签并将其克隆到自杀质粒pUT-Tn5Km2的转座子中,转化大肠杆菌S17-1λpir;然后用经转化的S17-1λpir与福氏志贺菌2a2457T交配,挑出对氨苄青霉素敏感、对卡那霉素和萘啶酮酸抗性的菌落。结果表明构建了包含4376个福氏志贺菌突变体信号标签诱变文库,为进一步鉴定该病原体的毒力基因打下了基础。To construct mutant library of Shigella flexneri2a by signature-tagged mutagenesis.Specific double strand DNA tags were obtained by PCR using the synthesized single strand DNAs as template,cloned into transpos on of the suicide plasmid pUTmini-Tn5Km2,and transformed into E.coli S17-1?pir;and then mating was carried out between the transformed S17-1?p ir cells and Shigella flexneri2a2457T(Nal r ),exconjugants were screened by picking nalidix-ic acid(Nal)resistant and kanamycin(Km)resistant but ampicillin(Ap)sensitive colonies.A mutant library of Shigella flexneri2a contained4376colonies was constructed by si gnature-tagged mutagenesis,This work provides the basis for identifying novel virulent genes of this pathogen.

关 键 词：专贺菌 福氏2A 信号标签 诱变 突变体文库 

分 类 号：R378.25[医药卫生—病原生物学] Q785[医药卫生—基础医学]