乳酸菌重组人碱性成纤维生长因子表达载体的构建及鉴定

Construction and identification of recombinant human basic fibroblast growth factor expression vector for lactic acid bacteria

目的构建乳酸菌重组人碱性成纤维蛋白表达载体,为口服治疗炎症性肠病探索新途径。方法利用融合PCR技术,将Ptuf启动子及pePN终止子与USP45-bFGF融合,构建全长片段后克隆于pbluescript载体中,经验证后亚克隆于pTRKH2载体中。结果双酶切及DNA测序结果证实成功构建乳酸菌表达载体pTR-bFGF。结论乳酸菌表达载体pTR-bFGF为实现bFGF在乳酸菌表达及口服治疗炎症性肠病奠定了基础。

Objective To explore a new way for the oral treatment of inflammatory bowel disease by constructing the recombinant human basic fibroblast protein expression vector. Methods The Ptuf promoter,pePN terminator, USP45 and bFGF were spliced by fusion PCR technology. The full-length constructed cDNA fragment was cloned into pBlueScript vector, as well as the pTRKH2 vector after identification. Results DNA sequencing and double enzyme digestion results showed that the expression vector pTR-bFGF was successfully constructed. Conclusion The pTR-bFGF plasmid laid a foundation for expression of bFGF in lactobacillus and oral treatment of inflammatory bowel disease.