猪霍乱沙门菌菌蜕的制备及免疫保护力研究

Preparation and immune efficacy of Salmonella choleraesuisghost

关键词:  猪霍乱沙门菌  菌蜕  免疫保护力  裂解基因E  裂解效率

Key words:  Salmonella choleraesuis  bacterial ghost  protective immunity  lysis gene E  lysis efficiency

摘要:

为制备猪霍乱沙门菌菌蜕并研究其免疫保护力,本实验克隆噬菌体phiX174裂解基因E,与pBV220连接,构建温控溶菌表达载体,将该载体转入猪霍乱沙门菌TTB1中,制备菌蜕并对其裂解率、安全性以及对小鼠的免疫保护力进行了研究。结果显示：成果克隆了裂解基因E、片段大小274bp,构建了温控溶菌质粒载体pBVE,制备了猪霍乱沙门菌菌蜕,其裂解率最高为99.46%、经冻干灭活残余活菌,安全性试验检测证实其安全后,对小鼠进行了免疫保护试验,其保护率为70%、与弗氏佐剂灭活苗保护率相当、优于甲醛灭活苗。研究结果为猪霍乱沙门菌引发疾病的防治奠定了基础。

Abstract:

To develop aSalmonella choleraesuis bacterial ghost vaccine and investigate its protective immunity.thisstudy, we cloned the lysis gene E from phage phiX174 andinserted into pBV220 vector ,which is a broad-host expression plasmid with temperature sensitive functions, and then transformed into Salmonella TTB1.We created the Salmonella choleraesuis ghostand studied its lytic efficiency and protective immunity. The results showed that the lysisgene E about 274bp was cloned, the temperature-induced lytic plasmid pBVE and Salmonella choleraesuis bacterial ghost were created.The lysis rate can be 99.46%and no life bacteria survived after freeze drying, The results of protective immunity showed that Salmonella ghost group providethe protection rate of 70%, The protection rate ofplasmid-type Salmonella ghost vaccine strain was comparable with that of Salmonella inactivated vaccine with adjuvant, which was better than that of the inactivated vaccine. The researchprovide a foundation for the prevention and treatment of diseases caused by Salmonella choleraesuis.