鸭沙门杆菌噬菌体尾部胞壁质水解酶基因克隆、表达及抗菌作用的试验研究  

Experimental studies of cloning,expression and antibacterial action of the murein hydrolase gene from the tail of bacteriophage for Salmonella anatis

作　　者：周蓝天 耿亚[1] 车晨[1] 刘福音[1] 郝葆青[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041

出　　处：《黑龙江畜牧兽医》2015年第10期30-33,38,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：西南民族大学大学生国家重点创新型科研项目(201410656017)

摘　　要：为了探索新型、特异性的鸭沙门杆菌噬菌体的杀菌作用及生物学特性,试验采用P22沙门杆菌噬菌体尾部区域编码胞壁质水解酶(GP4)的基因序列进行引物设计,以西南民族大学现代生物技术实验室前期分离的Ph1沙门杆菌噬菌体DNA为模板进行PCR扩增、克隆、重组和表达,并将获得的具有水解肽聚糖活性的重组蛋白(GP4-1,15 ku)进行体外抑菌试验和雏鸭沙门杆菌感染模型体内试验。结果表明:GP4-1抑制病原菌生长显著(P<0.05),可有针对性地杀灭鸭沙门杆菌临床分离株。说明其编码基因中除了含有胞壁质水解酶编码区域,还含有鸭沙门杆菌细胞壁表面特异性识别的结构域。To explore new and specific antibacterial action of bacteriophage for Salmonella anatis and its biological characteristics, the coding gene sequence of the gene 4 protein （ gp4 ） from the murein hydrolase in the tail of P22 baeterialphage for Salmonella anatis was adopted for the design of primers. The pre - separated DNA of Salmonellaphage Phi at the laboratory of modem biotechnology of Southwest University for Na- tionalities, was used as a template for the PCR amplification, gene cloning, recombination and expression. A recombinant protein （ GP4 - 1, 15 ku） with hydrolytic activity of peptidoglyean was successfully obtained, and then was used for an in vitro antibacterial test and in vivo test of ducking model of Salmonella anatis infection. The result showed that GP4 -1 could significantly inhibit （P 〈 O. 05 ） the growth of pathogenic bacteria, and was targeted to kill clinical isolates of Salmonella anatis. The results indicate that in addition to containing the coding region of murein hydrolase, its encoding genes of the GP4 - 1 also contain the structural domains for specific identification on the cell wall surface of Sal- monella anatis.

关 键 词：鸭沙门杆菌 噬菌体 胞壁质水解酶 基因克隆 表达 抗菌作用 

分 类 号：Q785[生物学—分子生物学]