维氏气单胞菌脂蛋白Lpp/LppB双基因缺失株的构建与验证  

Construction and identification of Aeromonas veronii lipoprotein Lpp/LppB double-gene deletion strain

作　　者：盛天鸽 宋格格 张俊辉[1] 王文东[1] 杨振国[1] 卢强[1] SHENG Tian-ge;SONG Ge-ge;ZHANG Jun-hui;WANG Wen-dong;YANG Zhen-guo;LU Qiang(Institute of Zoonosis,Jilin University,Changchun 130062,China)

机构地区：[1]吉林大学人兽共患病研究所,吉林长春130062

出　　处：《中国兽医科学》2021年第7期885-890,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31672686)。

摘　　要：在维氏气单胞菌单基因缺失株ΔLpp的基础上,通过同源重组方法获得脂蛋白LppB基因上、下游同源臂连接片段,依次构建重组克隆载体pMD18-T-L-UD1225和重组自杀性载体Pre112-L-UD1225,随后转入WM3064感受态细胞中作为供体菌,与受体菌ΔLpp进行结合转移,构建双基因缺失株ΔLpp/LppB,通过反转录验证并检测其遗传稳定性。结果显示,成功构建了遗传稳定性良好的双基因缺失株ΔLpp/LppB,其生长速率和生物被膜形成能力较野生株WT均下降。结果表明,协同缺失脂蛋白基因Lpp和LppB后,双基因缺失株ΔLpp/LppB的生长和生物被膜变化更显著,这有利于进一步推测脂蛋白基因的功能,为之后研究维氏气单胞菌的致病机制奠定了基础。Based on the single-gene deletion strain ΔLpp of Aeromonas veronii,the upper and downstream homologous arms connecting fragments of LppB gene of A.veronii were obtained by homologous recombination,and the recombinant cloning vector pMD18-T-L-UD1225 and the recombinant suicide carrier Pre112-L-UD1225 were constructed in sequence,and then transferred into WM3064 competent cells as donor bacteria,combined with the recipient bacteria ΔLpp,a double-gene deletion strain ΔLpp/LppB was constructed,and verification of its genetic stability was completed by reverse transcription.The results showed that a double-gene deletion strain ΔLpp/LppB with good genetic stability was successfully constructed,and its growth rate and biofilm formation ability were lower than that of wild strain WT.It showed that after the co-deleting lipoprotein genes Lpp and LppB,its growth and biofilm changes were more significant,which is conducive to further speculation of lipoprotein genes function.This result laid the foundation for the future study of the pathogenic mechanism of A.veronii.

关 键 词：维氏气单胞菌 Lpp基因 LppB基因 基因缺失 生物被膜 

分 类 号：S852.615[农业科学—基础兽医学]