单核细胞增生李斯特菌ncRNA rli60基因缺失株的构建研究  

Construction of Listeria monocytogenes ncRNA rli60 Gene Deletion Strain

作　　者：彭叶龙[1] 乔军[1] 孟庆玲[1] 谢堃[1] 赵海龙[1] 宋雪梅[1] 陈创夫[1] 

机构地区：[1]石河子大学动物科技学院动物疾病防控兵团重点实验室,石河子832003

出　　处：《石河子大学学报（自然科学版）》2014年第2期148-152,共5页Journal of Shihezi University(Natural Science)

基　　金：国家自然科学基金项目(31360596;30960274)

摘　　要：为了构建单核细胞增生李斯特菌(Listeria monocytogenes,LM)ncRNA rli60基因缺失株,采用基因重叠延伸PCR(SOE-PCR)的方法成功构建具有氯霉素抗性的pKSV7-Δrli60穿梭质粒,然后电转化至LM-SB5感受态细胞中,在温度和氯霉素的双重选择压力下进行同源重组,通过PCR进行重组菌鉴定。结果表明:成功筛选得到遗传性稳定的LM ncRNA rli60基因缺失株,为揭示LM ncRNA rli60基因功能提供了研究材料;同时,为进一步研究其在LM致病性及环境应激中的分子机制奠定了基础。To construct the ncRNA rli60 gene deletion strain of Listeria monocytogenes,the shuttle plasmid pKSV7-Δrli60 with chloramphenicol resistance was gained by overlap extension PCR successfully,which was electroporated into the LM-SB5 competent cell for homologous recombinant under the double pressure of temperature and chloramphenicol.Recombinant LM was identified by PCR.LM ncRNA rli60 gene deletion strain was successfully screened by PCR,which had stable genetic characteristic.This study provided materials for revealing the function of ncRNA rli60 gene and laid foundation for further study of the molecular mechanism in the LM pathogenicity and environmental stress.

关 键 词：单核细胞增生李斯特菌 NCRNA rli60基因 同源重组 重叠延伸PCR 

分 类 号：S852.61[农业科学—基础兽医学]