【作者】 张立强； 贺甜甜； 余少梅； 艾桃山； 喻运珍；

【Author】 Zhang liqiang;He tiantian;Yu shaomei;Ai taoshan;Yu yunzhen;Wuhan Fisheries Science Research Institution;Wuhan Chopper Fishery Bio-Tech Co.,Ltd.;

【机构】 武汉市水产科学研究所； 中博水产生物技术有限公司；

【摘要】 为了构建嗜水气单胞菌(Aeromonas Hydrophilia)AH-1菌株的平衡致死体系,实验通过Overlap PCR技术敲除了该菌的天冬氨酸半醛脱氢酶编码基因asd,结果显示该基因的敲除导致细菌对二氨基庚二酸(DAP)的强制性需求,本研究进一步构建了一个无抗生素元件的互补asd基因的质粒,该质粒可以互补asd基因的表达,将此质粒转入asd基因缺失突变株从而达到获得一株对此质粒严格需求的菌株,该菌株可以在没有DAP存在的条件下生存。研究表明本实验成功构建了嗜水气单胞菌平衡致死体系。更多还原

【Abstract】 In order to construct the balance lethal system of Aeromonas Hydrophilia AH-1 strain, this study deleted the Aspartate-Semialdehyde Dehydrogenase coding gene(asd) by Overlap PCR technology. The asd gene deletion resulted in a diaminopimelic acid(DAP)-dependent mutant. Furthermore, a complement plasmid with asd gene inserted in place of antibiotic-resistance was constructed. This plasmid expressed the asd gene and resulted the mutant DAP independent. This dependence on the Asd+ plasmid vector creates a balanced-lethal complementation between the bacterial strain and the recombinant plasmid. This study demonstrated a balanced-lethal system was constructed.更多还原