鳗弧菌菌蜕疫苗的初步制备  

Primarily Preparation of Vibrio anguillarum Ghost Vaccine

作　　者：刘丹阳 李超[1] 张敏[1] 姜光朋 周顺[1] LIU Danyang;LI Chao;ZHANG Min;JIANG Guangpeng;ZHOU Shun(College of Marine Science and Engineering,Qingdao Agricultural University,Qingdao 266200,China)

机构地区：[1]青岛农业大学海洋科学与工程学院,山东青岛266200

出　　处：《青岛农业大学学报（自然科学版）》2018年第4期316-321,共6页Journal of Qingdao Agricultural University(Natural Science)

基　　金：山东省重点研发计划(2016GSF115026)

摘　　要：菌蜕作为一种新型疫苗,较好的保留了细菌的原有形态及膜蛋白结构,能够诱导机体产生体液免疫反应及细胞免疫反应。为制备鳗弧菌菌蜕疫苗,本实验通过基因重组技术,构建了含噬菌体PhiX174裂解酶基因E(LysisE)及温控表达系统cIts857的重组载体pRK-857-E,通过结合转移技术,将重组表达载体转移至鳗弧菌,成功构建鳗弧菌菌蜕。结果表明,42℃升温诱导后,菌体OD600于1h时开始下降,3h后趋于平稳,表示细菌裂解基本完成。平板涂布结果显示,活菌数由诱导前的8.21×10~7 CFU/mL,下降至诱导后的2.31×10~3 CFU/mL,裂解效率接近99.999%。电镜对比观察诱导前后的细菌,发现诱导后的鳗弧菌形态未发生变化,可观察到直径为100~300nm的跨膜裂解孔洞,且有细胞质流出。本研究成功构建鳗弧菌菌蜕疫苗,并对其裂解效果进行检测,为弧菌菌蜕疫苗的开发奠定了基础。As a new vaccine, the bacterial ghosts preserve the original morphology of bacteria and the structure of membrane protein, which can induce the humoral immune response and cellular immune response. In order to prepare the Vibrioanguillarum bacterial ghost vaccine, a vector containing PhiX174 LysisE and temperature-controlling expression system cIts857 was constructed by gene recombination technology. The recombinant expression vector was transferred to V. anguillarum through the conjugal transfer technology, to successfully construct the V. anguillarum bacterial ghosts.The results showed that OD 600 began to decrease after 1 hour and became stable after 3 hours when induced temperature rose at 42°C, indicating that the bacterial lysis was basically complete. The spread plate results showed that the number of live bacteria count decreased from 8.21×10 7CFU/mL before induction to 2.31×10 3 CFU/mL after induction. The lysis efficiency was close to 99.999%. The morphology of bacteria showed no change by transmission electron microscopy observation after induction. However, transmembrane lysis pores with a diameter of 100~300 nm could be observed with cytoplasmic outflow. In the study, we successfully constructed the V. anguillarum bacterial ghosts and detected the lysis effect, which will provide the foundation for the development of Vibrio ghosts vaccine.

关 键 词：鳗弧菌 裂解 菌蜕 

分 类 号：Q939.91[生物学—微生物学]