1株黄牛源多源曼氏杆菌的分离鉴定与转铁结合蛋白B结构预测  

Isolation,identification and structural prediction of transferrin binding protein B in a Mannheimia varigena from cattle

作　　者：朱玉红 张疏桐 李志玲 徐雨蓓 杨雪赢 张贵东 韦国山 甘玲[1,2] 郭建华 ZHU Yuhong;ZHANG Shutong;LI Zhiling;XU Yubei;YANG Xueying;ZHANG Guidong;WEI Guoshan;GAN Ling;GUO Jianhua(College of Veterinary Medicine,Southwest University,Chongqing 402460,China;Immunology Research Center of Medicine Institute,Southwest University,Chongqing 402460,China)

机构地区：[1]西南大学动物医学学院,重庆402460 [2]西南大学医学研究院免疫学研究中心,重庆402460

出　　处：《中国兽医学报》2023年第4期706-712,共7页Chinese Journal of Veterinary Science

摘　　要：为确定引起肺炎病死黄牛的病原,从牛鼻拭子中分离纯化得到1株细菌,经生化试验和分子生物学方法鉴定为多源曼氏杆菌。利用生物信息学软件MEGA7.0对巴氏杆菌科细菌的16SrDNA和巴氏杆菌科、奈瑟菌科细菌的转铁结合蛋白B(TbpB)进行系统进化分析,并对TbpB进行了结构预测。结果显示,分离株16SrDNA基因序列与牛曼氏杆菌ZY190616株、肉芽肿曼氏杆菌ATCC49244株、多源曼氏杆菌10299株相似性最高,位于同一簇内,相似性超过97%;分离株TbpB序列与溶血曼氏杆菌、多源曼氏杆菌相似性较高,位于同一簇内,但与其他菌的TbpB序列相似性较低;TbpB结构模拟显示其序列与已经测定结构的溶血曼氏杆菌TbpB序列相似性较高。药敏试验显示该菌氨基糖苷类、β-内酰胺类抗生素有较强的耐药性,对头孢哌酮和氟喹诺酮类药物敏感小鼠致病性试验证实该分离菌株致死量约为2×10^(9) CFU。结果表明,该分离株为多源曼氏杆菌,其TbpB序列与溶血曼氏杆菌和多源曼氏杆菌相似性较高。本研究为后续毒力基因相关研究和疫苗开发提供了理论基础。To identify the pathogen from the cattle died of pneumonia,a strain of bacterium(RC2110)from nasal swab was isolated and purified.The bacterium was identified as Mannheimia varigena by biochemical tests and molecular biological methods.The 16SrDNA of Pasteurebacteriaceae and the transferrin binding protein B(TbpB)of Pasteurebacteriaceae and Neisseriaceae were analyzed by bioinformatics software MEGA 7.0.Then we predicted the structure of TbpB.The results showed that the sequence of 16SrDNA gene was highest similar to Mannheimia bovis strain ZY190616,Mannheimia granulomatis strain ATCC49244,Mannheimia varigena strain 10299.They belonged to the same cluster,and the similarity exceeded 97%.The TbpB sequence of the isolated strain was highly similar to that of Mannheimia bovis and Mannheimia varigena,and located in the same cluster.TbpB structure simulation showed that its sequence was similar to that of Mannheimia hemolytica.Drug sensitivity test showed that the strain had strong resistance to aminoglycoside andβ-lactam antibiotics,and pathogenicity test to cefoperazone and fluoroquinolone susceptible mice confirmed that the lethal dose of this strain was about 2×10^(9) CFU.The results showed that the TbpB sequence of the isolated strain was similar to that of Mannheimia hemolytica and Mannheimia varigena.This study provides a theoretical basis for subsequent virulence gene related research and vaccine development.

关 键 词：牛 多源曼氏杆菌 分离和鉴定 转铁蛋白结合蛋白B 结构预测和分析 

分 类 号：S852.61[农业科学—基础兽医学]