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菌蜕具有完整细菌表面抗原结构,可诱导机体的体液和细胞免疫应答,成为疫苗制备的候选之一,为了探讨鳗鲡迟缓爱德华氏菌菌蜕疫苗的可行性,实验采用基因重组技术构建了噬菌体PhiX174裂解酶基因(Lysis E)的温控表达载体,转化迟缓爱德华氏菌,成功制备其菌蜕,并对菌蜕形态、溶菌动力学、裂解效率以及制备条件等进行了研究。结果表明,细菌菌蜕表面形成溶菌孔道,细胞因内容物流失而发生明显的皱缩;构建的迟缓爱德华氏菌诱导后1 h开始裂解,5 h后裂解基本完成,裂解效率为99.99%,冷冻干燥后重悬涂布平板,未检出活菌,电镜观察表明冻干前后细胞形态未见明显变化;构建的迟缓爱德华氏菌分别在OD600值为0.4和0.6进行诱导,其裂解过程和裂解效率没有明显区别;分别用LB、BHI、NB 3种培养基进行比较研究,其中LB培养基制备的菌蜕细胞较完整、裂解完全,是制备迟缓爱德华氏菌菌蜕最优培养基。本研究成功构建了鳗源迟缓爱德华氏菌菌蜕,并对其制备条件进行了优化,为鳗鲡爱德华氏菌病疫苗开发奠定了基础。
Bacterial slough has complete bacterial surface antigen structure, which can induce humoral and cellular immune responses of the body and become one of the candidates for vaccine preparation. In order to investigate the feasibility of slow-release Erigeron residenciae vaccine, the recombinant phage PhiX174 Lysis E gene was transformed into E.coli and transformed into E.coli. The bacterium shedding was successfully prepared. The morphology, bacteriolytic kinetics, pyrolysis efficiency and preparation conditions were studied. The results showed that the bacterial cells formed lysis holes on the surface of the bacteria, and the cells collapsed obviously due to the loss of the contents. The lysis of the constructed R. edwardsii lysis started 1 h after induction, and the lysis was basically completed after 5 h. The lysis efficiency was 99.99% After drying, the plates were resuspended and the viable cells were not detected. Electron microscopy showed no significant changes in cell morphology before and after lyophilization. The constructed E. canadensis strains were induced at OD600 values of 0.4 and 0.6, respectively. The lysis and lysis efficiency No significant difference; respectively LB, BHI, NB three kinds of media for comparative studies, including LB medium prepared cells more complete cell lysate, complete cleavage, is the preparation of the slow Edwardsiella bacteria optimum shedding medium. In this study, we successfully constructed a slow-release Eeliana edulis bacterium, and optimized its preparation conditions, laying a foundation for the development of vaccination of Edwardsiella eel disease.