肠炎沙门菌sopE2基因缺失株的构建及其生物学活性的鉴定

发布时间：2018-01-29 20:13

本文关键词： 肠炎沙门菌 sopE 基因缺失生物学活性鉴定　出处：《中国兽医科学》2017年03期 　论文类型：期刊论文

【摘要】：为研究sopE 2基因对肠炎沙门菌(Salm onella enteritidis)的生物学特性的影响及其与sopE功能差异,本研究以λ-Red介导的同源重组法构建了肠炎沙门菌C50336ΔsopE 2缺失株,并构建了以p BR322质粒介导的sopE2基因回复株。测定了肠炎沙门菌野生株、sopE 2缺失株及其回复株的基本生物学特性,包括对人肠上皮细胞Caco-2 BBE及小鼠巨噬细胞RAW 264.7的黏附、侵袭能力及对小鼠的半数致死量。结果显示,肠炎沙门菌sopE 2基因缺失对其生长、生化特性无影响;肠炎沙门菌C50336对Caco-2 BBE细胞的黏附率约为20%,侵袭率约为10%,对RAW264.7细胞的黏附率约为55%,侵袭率约为20%,sopE 2基因缺失对肠炎沙门菌C50336黏附能力无影响,侵袭能力有所下降,但无统计学差异;肠炎沙门菌C50336对小鼠的半数致死量为5.0×105CF U,sopE 2基因缺失株及回复株对小鼠的半数致死量均为3.2×105 CFU,表明sopE 2基因缺失对肠炎沙门菌的毒力无显著影响。本研究为进一步探讨sopE 2基因在肠炎沙门菌感染过程中的功能奠定了基础。
[Abstract]:To study the effect of sopE 2 gene on the biological characteristics of Salm onella enteritidissis and its functional difference with sopE. In this study, Salmonella enteritidis C50336 螖 sopE 2 deletion strain was constructed by homologous recombination mediated by 位 -Red. The sopE2 gene response strain mediated by p BR322 plasmid was constructed, and the basic biological characteristics of the wild Salmonella enteritidis strain were determined. The adhesion, invasiveness and lethal dose to human intestinal epithelial cells (Caco-2 BBE) and mouse macrophages (RAW 264.7) were included. The absence of sopE 2 gene had no effect on the growth and biochemical characteristics of Salmonella enteritidis. The adhesion rate of Salmonella enteritidis C50336 to Caco-2 BBE cells was about 20, the invasion rate was about 10 and the adhesion rate to RAW264.7 cells was about 55%. The invasion rate was about 20% and the loss of sopE2 gene had no effect on the adhesion ability of Salmonella enteritidis C50336, but the invasive ability was decreased, but there was no statistical difference. The lethal dose of Salmonella enteritidis C50336 to mice was 5.0 脳 105 CF U. The median lethal dose of sopE 2 gene deletion strain and recovery strain to mice was 3.2 脳 10 5 CFU. The results showed that the absence of sopE 2 gene had no significant effect on the virulence of Salmonella enteritis. This study laid a foundation for further study of the function of sopE 2 gene in the process of Salmonella enteritis infection.
【作者单位】：扬州大学江苏省人兽共患病学重点实验室江苏省动物重要疫病与人兽共患病防控协同创新中心;
【基金】：国家自然科学基金项目(31320103907) 江苏省高校研究生科研创新计划项目(KYZZ_0368)
【分类号】：S852.61
【正文快照】： 沙门菌为革兰阴性、兼性厌氧胞内寄生菌,可感染包括人在内的多种宿主。沙门菌感染已成为一种重要的公共安全问题[1],在美国,沙门菌感染每年导致超过100万人致病及约350人死亡[2]。在世界范围内,沙门菌感染每年大约导致938万人致病,155 000人死亡[3]。在过去20年中,尽管有各种

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