目的:构建连接乳酸菌胞外蛋白水解酶基因的非抗性重组质粒。方法:以瑞士乳杆菌基因组DNA为模板,克隆胞外蛋白水解酶基因,连接到以thyA为选择压力的非抗性穿梭表达载体pW425et上,构建重组质粒pW425et-R,转化入thyA基因缺陷型E.coliX13感受态细胞,SDS-PAGE电泳检测显示该水解酶得到了表达。结果:成功构建了重组质粒pW425et-R,胞外蛋白水解酶基因在E.coliX13中得到正确表达。结论:成功地表达了乳酸菌胞外蛋白水解酶基因,可为进一步制备具降血压功能基因工程乳酸菌提供参考。
Objective:To construct a recombinant plasmid carrying extracellular protease gene from Lactobacillus.Methods:The extracellular protease gene was cloned from genomic DNA of L.helveticus and inserted into the expression vector pW425et.Then the recombination plasmid was transformed into the competence thyA gene-mutant E.coli X13.The expressed protein was analyzed by SDS-PAGE.Results:The recombinant shuttle plasmid pW425et-R was constructed successfully and the Lactobacillus extracellular proteinase gene was expressed in E.coli X13.Conclusion:Our research idea is feasible for the successful expression of Lactobacillus extracellular protease gene in E.coli X13,which will provide a basis for further preparation of recombinant Lactobacillus with anti-hypertensive function.