基因工程技术制备梅毒螺旋体重组抗原的研究

Study on Preparation of Recombinant Treponema Pallidum Antigen with Genetic Engineering Technique

目的 用基因工程技术制备梅毒螺旋体特异蛋白抗原，以解决梅毒螺旋体不能体外培养而难以获得足量的纯梅毒螺旋体特异蛋白抗原的难题。 方法 以 PCR技术克隆出目的基因，重组后通过 DNA序列测定验证重组质粒中连结有目的基因片段；以 pMALTM-c2为质粒载体， TB1大肠杆菌为宿主菌进行目的基因的转化、诱导和表达，并对表达产物进行蛋白印迹试验检测其血清学活性。结果 PCR克隆出梅毒螺旋体 15 000、 17 000以及 47 000蛋白的基因克隆，其中梅毒螺旋体 17 000以及 47 000在 TB1大肠杆菌中得到稳定的表达，且表达产物显示与梅毒患者血清有非常特异的血清学反应。结论 梅毒螺旋体之目的基因在大肠杆菌中得到了成功表达，并具特异的血清学活性，从而为建立国产化梅毒血清学诊断的新方法奠定了基础。

Objective To prepare specific recombinant Treponema pallidum antigen with genetic engineering technique. Methods The specific antigen genes were cloned by polymerase chain reaction (PCR) technique, and checked by sequencing. The procedures of translation, induction and expression were performed using pMALTM-c2. Results The genes encoding TP 15 000, TP 17 000 and TP 47 000 were obtained by PCR. The antigens of TP 17 000 and TP 47 000 were successfully expressed in E.coli TB1. The recombinant TP 17 000 and TP 47 000 produced showed a highly specific serological activity. Conclusion The specific TP antigen gene is successfully expressed in E.coli with products showing high specificity to the sera of patients with syphilis, which might be used for the diagnosis of syphilis.