梅毒螺旋体TpN15重组抗原的克隆与表达

Cloning and expression of recombinant treponema pallidum TpN15

目的：用基因工程技术表达梅毒螺旋体TpN15重组抗原，为进一步研制梅毒螺旋体疫苗和ELISA诊断试剂盒奠定基础。方法：采用PCR技术扩增梅毒螺旋体TpN15基因，进行T-A克隆及测序，然后亚克隆到原核表达载体中，以亲和层析法纯化重组蛋白。结果：成功地构建了pET32-TpN15重组表达载体，TpN15重组蛋白在大肠杆菌BL21中得到稳定表达。结论：梅毒螺旋体TpN15基因在大肠杆菌中的成功表达为临床检测梅毒感染的新方法和梅毒螺旋体疫苗的研制奠定了基础。

Objective To produce recombinant TpN15 treponema pallidum antigen with genetic engineering technique, so as to set basis for the preparation of ELISA diagnostic kit and TP vaccines. Methods The TpN15 gene was amplified from TP genome by PCR technique. Amplification fragments of the target genes were sequenced after T-A cloning and cloned into prokaryotic expression vector to express recombinant TpN15 protein. The recombinant fusion antigen was purified with affinity chromatography. Results The expression vector pET32-pN15 was constructed successfully, and TpN15 protein was highly expressed in E. coli BL21. Conclusion The successful acquirement of recombinant TpN15 protein provides a basis for establishment of new clinical diagnosis method of syphilis and preparation of TP vaccines.