鸭疫里默菌aroA基因缺失突变株的构建及生物学特性研究  

Construction and Characterization of R.anatipestifer aroA Deletion Strain

作　　者：张硕[1] 刘宁[1] 卢媞 王颖[1] 周祖涛[1,2] 刘梅[1,2] 胡思顺[1,2] 栗绍文[1,2] 毕丁仁[1,2] 李自力[1,2] 

机构地区：[1]华中农业大学动物医学院,湖北武汉430070 [2]农业微生物学国家重点实验室,湖北武汉430070

出　　处：《动物医学进展》2015年第12期1-5,共5页Progress In Veterinary Medicine

基　　金：武汉市科技计划项目(2015020101010070)

摘　　要：构建鸭疫里默菌aroA基因缺失株,并对其生物学特性及毒力进行研究。以本实验室分离的鸭疫里默菌YM株(RA-YM)为亲本株,建立pRE112-LSR重组自杀性质粒,通过结合转移的方法缺失RAYM株aroA基因,并利用PCR方法对突变株进行鉴定,成功构建了鸭疫里默菌aroA基因缺失株。生物学特性检测结果显示,鸭疫里默菌aroA基因缺失株不再水解精氨酸;体外生长曲线对数期细菌生长速度减缓,在稳定期趋于一致;半数致死量与野生株相比下降了3倍左右;aroA基因缺失株在血液、心、肝、脑等组织的载菌量与亲本株相比有所下降,但差异不显著。研究结果为深入研究鸭疫里默菌的分子致病机理和研制基因工程疫苗奠定了良好的基础。R.anatipestifer aroA deletion strain was constructed and its biological characteristics and virulence were detected.Suicide recombinant plasmid of pRE112-LSR was constructed,and the aroA gene in R.anatipestifer YM strains was successfully deleted by conjugation and identified by PCR.The results showed that the main biological characteristics were similar with the wild-type strain,while the aroA mutant could not hydrolyze arginine;Growth rate of the aroA mutant in vitro was slower than wild-type strain within logarithmic phase,and similar to the wild-type strain at stable phase;LD50of the aroA mutant was decreased three times than that of wild-type strain;Quantity of isolated bacteria from ducking tissues showed that the aroA mutant was less than it in blood,heart,liver and brain of wild-type strain,but it was not significant.These results provided a good foundation for the further study of the molecular pathogenesis of R.anatipestifer and development of genetic engineering vaccines.

关 键 词：鸭疫里默菌 aroA基因 基因缺失 生物学特性 毒力因子 

分 类 号：S852.61[农业科学—基础兽医学]