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目的 为构建霍乱弧菌口服活疫苗提供稳定实用的载体系统。方法 在改良MM基本培养基的基础上 ,建立了霍乱弧菌thyA基因突变菌株的筛选方法 ,继而用PCR插入法克隆了大肠杆菌thyA基因全序列 (pXXB10 6 ) ,并电击转化入霍乱弧菌thyA基因缺陷株 (PL10 2 ) ,使后者在MM改良培养基上的生长恢复为野生状态。结果 选育出的thyA基因突变的霍乱弧菌IEM10 1菌株PL10 2突变性状及生物学性状基本稳定。进而构建了以thyA基因为选择压力的霍乱弧菌染色体 质粒致死平衡系统。结论 构建成的抗原呈递系统作为构建肠道疫苗的受体菌株是稳定的 ,也是适用的。
Objective To provide a stable and practical vector system for the construction of oral live Vibrio cholerae vaccine. Methods Based on the modified MM basic medium, a screening method was established for the mutant strain of Vibrio cholera thyA. The full-length sequence of the thyA gene of E. coli was cloned by PCR (pXXB10 6) and transformed into Vibrio cholerae thyA The gene-deficient strain (PL10 2) restored the latter to a wild state on the MM modified medium. Results The mutation and biological characteristics of PL10 2 strain of Vibrio cholerae IEM10 1 with mutant thyA mutation were basically stable. Then we constructed a lethal balance system of cholera chromosome plasmid with thyA gene as selective pressure. CONCLUSION: The constructed antigen presenting system is stable and suitable as a recipient strain for constructing an intestinal vaccine.