肠炎沙门菌非编码小RNA RyhB-1和IsrE对清远麻鸡的致病性  

Pathogenicity of Salmonella enteritidis non-coding small RNAs RyhB-1 and IsrE in Qingyuan chickens

作　　者：倪杰[1,2] 孟宪臣[1] 李芙蓉[3] 谢静[3] 陆广富[3] 朱春红[4] 陈凯[1,2] 孟霞[1,2] 朱国强[1,2] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009 [3]江苏农牧科技职业学院,江苏泰州225300 [4]江苏省家禽科学研究所,江苏扬州225125

出　　处：《中国兽医学报》2017年第9期1687-1692,共6页Chinese Journal of Veterinary Science

基　　金：江苏省高校自然科学研究资助项目(14KJB230002);兽医生物技术国家重点实验室开放课题资助项目(SKLVBF201509);扬州市自然科学基金青年科技人才资助项目(YZ2014019);科技部畜禽重大疫病防控与高效安全养殖综合技术研发重点专项资助项目(2016YFD0500900);江苏高校优势学科建设工程资助项目(PAPD);国家自然科学基金资助项目(31101826;31270171);泰州市农业科技发展资助项目(TZ201422)

摘　　要：RyhB是一种大小为90bp左右的细菌非编码小RNA,已有研究表明存在于鼠伤寒沙门菌和霍乱弧菌中的2种RyhB同源物RyhB-1和IsrE可调控细菌生物膜形成、趋化性和耐酸性。为研究RyhB在肠炎沙门菌致病性上的作用,以及2种同源物能否对毒力调控发挥协同作用,本试验利用λ-Red同源重组系统构建了ryhB-1和isrE单、双缺失株,利用表达载体pBR322和pACYC184分别构建了回补质粒并导入缺失株50336△ryhB-1,50336△isrE,50336△ryhB-1/△isrE中,获得各突变株相应的回补菌株,检测了野生株和各突变株对1日龄雏鸡半数致死量(LD50)、致死率、体内分布等致病性差异。结果发现,ryhB-1和isrE基因缺失后导致肠炎沙门菌毒力减弱,双基因缺失株毒力下降更为明显,表明2种小RNA均对肠炎沙门菌致病性具有增强作用,且二者对毒力的调控具有协同作用。RyhB is a kind of about 90 bp bacterial non-coding small RNA. It was reported that two RyhB homologs RyhB-1 and IsrE,whieb existed in the Salmonella typhirnurium and Vibrio chol- erae could regulate biofilm formation, chemotaxis and acid resistance. To study the pathogenicity and cooperative function of RyhB-1 and IsrE, we constructed ryhB-1 and isrE single and double deletion mutants by the X-Red-based recombination system and corresponding complemented mu- tants by constructing complemented plasmid with pBR322 and pACYC184,and transferring to de- letion mutants 50336△ryhB-1,50336△isrE and 50336△ryhB-1/△isrE. Then we tested the path- ogenic differences of wild type,ryhB-1 and isrE deletion mutants and complemented mutants by inoculating above strains to 1-day-old chickens and detecting LD56, mortality rate and bacterial dis- tribution in chickens. The results showed that the virulence of Salrnonella enteritidis was de- creased in the ryhB-1 or isrE gene deletion strain,especially in ryhB-1 and isrE double deletionstrain. It indicated that ryhB-1 and isrE gene could enhance pathogenicity of Salmonella enteriti dis. The two sRNAs have a cooperative effect in virulence regulation.

关 键 词：肠炎沙门菌 ryhB-1 ISRE 半数致死量 致病性 

分 类 号：S852.62[农业科学—基础兽医学] R535[农业科学—兽医学]