hmpA基因对鸡白痢沙门菌在巨噬细胞内生存影响的研究  

Survival influence of hmpA gene on Salmonella pullorum in macrophages

作　　者：潘明星 康喜龙 周莹莹 孟闯 潘志明 焦新安 PAN Ming-xing;KANG Xi-long;ZHOU Ying-ying;MENG Chuang;PAN Zhi-ming;JIAO Xin-an(Jiangsu Key Laboratory of Zoonosis/Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonosis/Key Laboratory of Prevention and Control of Biological Hazard Factors(Animal Origin)far Agrifbod Scfety and Quality,Ministry of Agriculture/Joint International Research Lab oratory of Agriculture and Agriproduct Safety of the Ministry of Education,Yangzhou University Hangzhou 225009,China)

机构地区：[1]扬州大学江苏省人兽共患病学重点实验室江苏高校动物重要疫病与人兽共患病防控协同创新中心农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室农业与农产品安全国际合作联合实验室,江苏扬州225009

出　　处：《中国兽医科学》2022年第1期104-110,共7页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31730094,31902278);江苏省政策引导类计划项目(BZ2020013);中国博士后基金面上项目(2018M642333)。

摘　　要：为了探究hmpA基因对鸡白痢沙门菌在胞内生存的影响,本研究利用同源重组方法构建了鸡白痢沙门菌449/87的hmpA基因缺失株(449/87ΔhmpA),同时利用原核表达载体pMMB207构建了基因回复株449/87ΔhmpA::hmpA,并对其生长曲线、生化特性以及其感染鸡源巨噬细胞HD11后一氧化氮合酶(NOS)活性、活性氧(ROS)和胞内生存情况进行测定。结果显示,hmpA基因的缺失对鸡白痢沙门菌生长速度和生化特性无影响。与野生株和回复株相比,449/87ΔhmpA感染巨噬细胞后,NO、NOS活性和ROS显著升高,胞内细菌载量明显减少,表明hmp A基因的缺失能够增强鸡白痢沙门菌诱导巨噬细胞的NOS活性和ROS的生成,降低细菌在胞内的存活能力。本研究为进一步探究hmpA基因在鸡白痢沙门菌感染过程中的功能奠定了基础。In order to study the influence of Salmonella pullorum hmp A gene on the bacteria survival in the macrophages,the hmpA gene deletion strain of Salmonella pullorum 449/87(449/87ΔhmpA) was implemented through a homologous recombination method.Moreover,the complementation strain(449/87Δhmp A::hmp A) was constructed via the prokaryotic expression vector p MMB207.The growth curve and biochemical characteristics of 449/87Δhmp A were determined.The intracellular survival of S.pullorum,as well as nitric oxide synthase(NOS) activity,and reactive oxygen species(ROS) generation in chicken-derived macrophages HD11 were evaluated after Salmonella infection.All results indicated that the deletion of hmp A gene had no obvious effect on the growth rate and biochemical characteristics of S.pullorum.Nevertheless,NO release,ROS generation and NOS activity of macrophages HD11 increased,and the amount of intracellular bacteria were significantly reduced after 449/87Δhmp A infection,compared with parental 449/87 and 449/87Δhmp A::hmp A.This study demonstrated that the deletion of hmp A gene could induce NOS activity and produce more ROS in macrophages,and weaken the intracellular survival ability of S.pullorum,which would lay an important foundation for further research on the function of the hmp A gene during S.pullorum infection.

关 键 词：鸡白痢沙门菌 hmpA基因 一氧化氮合酶活性 活性氧 胞内增殖 

分 类 号：S852.612[农业科学—基础兽医学]