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目的探讨利用噬菌体PhiX174 E基因制备人伤寒沙门菌Ty21a菌蜕的可行性以及适当制备条件。方法将PhiX174 E基因克隆至温控表达系统,筛选获得裂解效率较高的质粒pMBE,通过优化转化方法将pMBE导入Ty21a,利用温控表达系统进行Ty21a菌蜕制备。菌落计数计算菌蜕裂解效率,并通过扫描电镜和透射电镜观察其形态结构。结果裂解质粒pMBE介导大肠杆菌DH5α的裂解效率达99.99%;通过电击转化方法获得重组Ty21a(pMBE),利用温控表达系统成功制备Ty21a菌蜕,裂解效率达96.77%。电镜观察菌蜕结构完整并呈空泡状结构,并能看到溶菌孔道。结论 PhiX174 E基因表达可实现人伤寒沙门菌Ty21a裂解形成Ty21a菌蜕,为其作为疫苗和药物递送载体的研究奠定了基础。
Objective To investigate the feasibility and preparation of Salmonella typhi Ty21a by phage display of PhiX174 E gene. Methods PhiX174 E gene was cloned into a temperature-controlled expression system. The plasmid pMBE with high cleavage efficiency was screened. The pMBE was introduced into Ty21a by the optimized transformation method, and the Ty21a strain was prepared by temperature-controlled expression system. The colony counts were used to calculate the efficiency of bacterial decolourization and lysis, and the morphological structure was observed by scanning electron microscopy and transmission electron microscopy. Results The lytic efficiency of E. coli DH5α was 99.99%. The recombinant Ty21a (pMBE) was obtained by electroporation and the Tyha bacterium was successfully prepared by temperature-controlled expression system. The efficiency of cleavage was 96.77%. Electron microscopy bacterial spore structure intact and vacuolar structure, and can see the lysozyme channel. Conclusion PhiX174E gene expression can be achieved Tyrosinase Ty21a cleavage to Ty21a bacteria shed, as its vaccine and drug delivery vector lays the foundation.