鸭沙门菌rfaH基因缺失株的构建及其生物学特性研究  

Construction of the rfaH Gene Deletion Strain in Salmonella anatum and Study of Its Biological Characteristics

作　　者：宋亚雯 高宇杰 张晓荟 王温馨 葛同鑫 翟瑞东 宋厚辉 程昌勇 李爱云[2] 韩月 SONG Ya-Wen;GAO Yu-Jie;ZHANG Xiao-Hui;WANG Wen-Xin;GE Tong-Xin;ZHAI Rui-Dong;SONG Hou-Hui;CHENG Chang-Yong;LI Ai-Yun;HAN Yue(College of Animal Science and Technology/College of Veterinary Medicine/Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province/Zhejiang Provincial Engineering Research Center for Animal Health Diagnostics/Advanced Technology,Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management/China-Australia Joint Laboratory for Animal Health Big Data Analytics,Zhejiang A&F University,Hangzhou 311300,China;Women's Hospital School of Medicine Zhejiang University,Hangzhou 310006,China)

机构地区：[1]浙江农林大学动物科技学院/动物医学院/浙江省畜禽绿色生态健康养殖应用技术研究重点实验室/动物健康互联网检测技术浙江省工程研究中心/浙江省动物医学与健康管理国际科技合作基地/中澳动物健康大数据分析联合实验室,杭州311300 [2]浙江大学医学院附属妇产科医院,杭州310006

出　　处：《农业生物技术学报》2025年第3期642-649,共8页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划项目(2023YFD1801000);浙江省自然科学基金(LQ24C010005);国家自然科学基金(32102671l31902280)。

摘　　要：鸭沙门菌(Salmonella anatum)是引起鸭沙门菌病的主要病原微生物。rfaH是编码抗转录终止因子RfaH的基因,在细菌转录、感染等生物学特性中具有重要作用。为了研究rfaH基因对鸭沙门菌生长、运动、生物被膜和感染细胞等生物学特性的影响,本研究通过Red同源重组技术,利用鸭沙门菌分离株SA01构建了rfaH基因缺失株ΔrfaH和回补株CΔrfaH。测序结果表明,SA01的rfbH基因缺失株ΔrfaH和回补株CΔrfaH构建成功。rfbH基因缺失后,SA01生长速率不变,但O抗原合成受损,细菌运动能力显著降低,生物被膜形成能力增强,对鸡胚成纤维细胞DF-1细胞的黏附和侵袭能力增强,在小鼠单核巨噬细胞RAW264.7中的增殖能力减弱。研究结果表明,rfbH基因参与沙门菌的运动、生物被膜形成和感染细胞等生物学过程。本研究初步阐明了SA01的rfaH基因的生物学功能,为开发基于鸭沙门菌的递呈载体疫苗提供了理论依据。Salmonella anatum is a major pathogen that causes salmonellosis in ducks(Anatidae).rfaH encodes the transcriptional anti-termination factor RfaH,which is essential for bacterial transcription,infection,and other biological functions.This study investigated the effect of the rfaH gene on various biological aspects of S.anatum,including growth,motility,biofilm formation,and cell infection.To deeply understand the biological roles of the rfaH gene in S.anatum,an rfaH gene deletion strainΔrfaH and the complement strain CΔrfaH derived from the S.anatum isolate strain SA01 were generated using Red homologous recombination technology.The sequencing results showed that the rfaH gene deletion strainΔrfaH and the complement strain CΔrfaH of SA01 were successfully constructed.After the deletion of the rfbH gene,the growth rate remained unchanged,yet the O-antigen synthesis was impaired,leading to a significant decrease in bacterial motility,an enhancement in biofilm formation,an increased adhesion and invasion capacity against DF-1 cells,and a weakened proliferation ability in RAW264.7 cells.The findings indicated that the rfaH gene was involved in bacterial motility,biofilm formation,and cell infection.This study establishes a solid platform for future research on the biological functions of the rfaH gene and the development of Salmonella-based delivery vectors for vaccine applications.

关 键 词：鸭沙门菌 rfaH基因 生物学特性 

分 类 号：S855[农业科学—临床兽医学]