CAJ | 학술논문

目的：：在大肠杆菌中重组表达梅毒螺旋体TpN17抗原，通过亲和层析方法进行纯化，获得纯度较高的TpN17抗原，利用该抗原建立梅毒诊断血清学方法。方法：将化学合成的TpN17基因序列，克隆到pET 22b质粒并转化到大肠杆菌BL21菌株中进行IPTG的诱导表达。利用Ni2+亲和层析柱对表达抗原进行纯化。结果：对插入片段进行测序，证实该序列结果与合成序列一致，并在大肠杆菌中成功表达TpN17抗原，通过亲和层析获得纯度较高的目的抗原。结论：本研究中构建的大肠杆菌重组表达载体可有效的表达TpN17抗原，经纯化后的抗原，可有望进一步应用于梅毒感染的血清学检测试剂盒的研发。
목적：：재대장간균중중조표체매독라선체TpN17항원，통과친화층석방법진행순화，획득순도교고적TpN17항원，이용해항원건립매독진단혈청학방법。방법：장화학합성적TpN17기인서렬，극륭도pET 22b질립병전화도대장간균BL21균주중진행IPTG적유도표체。이용Ni2+친화층석주대표체항원진행순화。결과：대삽입편단진행측서，증실해서렬결과여합성서렬일치，병재대장간균중성공표체TpN17항원，통과친화층석획득순도교고적목적항원。결론：본연구중구건적대장간균중조표체재체가유효적표체TpN17항원，경순화후적항원，가유망진일보응용우매독감염적혈청학검측시제합적연발。
Objective: To express and purify the treponema pallidum 17 (TpN-17) protein by using a prokaryotic expression vector, and to establish a serological diagnosis method of syphilis. Methods: The TpN17 gene was synthesized and cloned into the pET 22b vector. Then the TpN17 protein was induced to express with IPTG and to be purified through affinity chromatography method. Results: The sequencing result of TpN17 DNA was confirmed in accordance with our expectation. And the protein was successfully expressed and purified. Conclusion: The vector can effectively express the TpN17 protein and the purified antibodies can be applied to develop the EIA detection kits of syphilis.