梅毒螺旋体0751基因的克隆、表达及鉴定

Cloning,expression and identification of the 0751 gene from Treponema pallidun

目的构建梅毒螺旋体0751基因的原核表达质粒,诱导重组蛋白表达及进行免疫学鉴定。方法以梅毒螺旋体基因组DNA为模板,PCR扩增梅毒螺旋体0751基因,构建其重组原核表达载体,并采用Western-blot方法初步鉴定该蛋白的特异性。结果成功扩增出梅毒螺旋体0751基因,该基因在大肠杆菌表达系统中得到表达,并且特异性识别梅毒患者血清。结论在大肠杆菌中成功表达梅毒螺旋体0751基因,为梅毒螺旋体的诊断提供了新的特异性抗原。

Objective The aim of the study was to clone,express and identify of 0751 gene from Treponema pallidun. Methods 0751 gene was amplified by the PCR from the genomic DNA of Treponemapallidun and cloned into PET-28a （＋） vector.After induction by IPTG,the 0751 protein was identified by Western-blot. Results The 0751 gene （552bp）was amplified and the recombinant protein was expressed successfully.Western-blot showed that recombinant 0751 could be identified specifically by Treponema pallidun infected serum. Conclusion The recombinant 0751 protein was expressed successfully and contributed to the diagnosis of Treponemapallidun infection.