【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。更多还原

【Abstract】 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.更多还原