hok/sok-parDE增强鼠伤寒沙门菌菌毛编码质粒稳定性研究  

The Combination of hok/sok-parDE Enhance Stability of the Plasmid Encoding Salmonella Type Ⅰ Fimbrial Gene

作　　者：杨维红[1] 陈杨[1] 张成栋[1] 孙国忠[1] 凌洁玉[1] 涂玲玲[1] 陈焕春[1] 郭爱珍[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：《动物医学进展》2007年第9期7-11,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(30571386)

摘　　要：将编码两个质粒稳定系统的融合基因克隆至表达鼠沙门菌Ⅰ型菌毛(fim)基因的质粒pAZ44中,获得pAZ44-h。将两质粒分别转化鼠伤寒沙门菌,连续培养144 h,每隔12 h转接1代,并取样,用抗性平板计数细菌,通过抗性检测质粒存在状况。结果发现,质粒pAZ44在细菌培养12 h^24 h后开始丢失,而含稳定系统的质粒pAZ44-h在连续培养10代或120 h都未发生丢失。表明aphA-hok-parDE序列显著增强了质粒的稳定性,为进一步研究鼠沙门菌Ⅰ型菌毛的功能奠定了基础。In order to enhance plasmid stability, the fusion of hok/sok and parDE fragments （aphA-hokparDE ）of R1 and RK2 was subcloned into pAZ44 to obtain pAZ44-h. The plasmids pAZ44 and pAZ44-h were respectively transformed into Salmonella typhimurium by electroporation. The single colonies were picked up and cultured in LB broth. For every 12 h, 10 ml of the samples was transferred to a new flask until 144 h. At the same time, the bacteria were inoculated onto the LB plates with different antibiotics. By comparing the bacterial number, the existence of plasmids was detected. The results showed that the plasmid pAZ44 without the stabilization system began to loss plasmid at the 12 h-24 h of culture, while pAZ44-h did not lose the plasmid during the first 10 passages （or 120h of culture）. It demonstrated that aphA-hok-parDE previously enhanced the plasmid stability. This finding laid a good foundation for further study of Salmonella typhimurium type Ⅰ fimbriae.

关 键 词：质粒稳定性 hok/sok parDE 沙门菌 

分 类 号：S852.612[农业科学—基础兽医学]