论文部分内容阅读
目的在原核系统内获得结核分枝杆菌Rv3425蛋白可溶性表达并进行纯化,Western-blot技术初步鉴定重组蛋白的抗原性和特异性。方法将Rv3425核酸编码序列克隆至融合表达载体p ET-Dsb C中,然后进行融合蛋白Dsb CRv3425表达并实现亲和纯化,用Western-blot分析其抗原性和特异性。结果 Rv3425基因在大肠杆菌中获得可溶性表达,亲和纯化的Dsb C-Rv3425蛋白与结核病患者血清标本呈强阳性反应,与健康人血清标本呈阴性反应。结论Dsb C-Rv3425蛋白在大肠杆菌中以可溶性表达形式存在,具有较好的抗原特异性和免疫原性,对结核病诊断有潜在的应用价值。
Objective To obtain the soluble expression of Mycobacterium tuberculosis Rv3425 protein in prokaryotic system and to purify it. Western-blot technique was used to identify the antigenicity and specificity of the recombinant protein. Methods The coding sequence of Rv3425 was cloned into the fusion expression vector p ET-Dsb C, then the fusion protein Dsb CRv3425 was expressed and affinity-purified, and its antigenicity and specificity were analyzed by Western-blot. Results The Rv3425 gene was expressed in Escherichia coli. The affinity purified Dsb C-Rv3425 protein was strongly positive to the serum of patients with tuberculosis and negative to the serum of healthy people. Conclusion The Dsb C-Rv3425 protein exists in soluble form in E. coli and has good antigen specificity and immunogenicity. It has potential value in the diagnosis of tuberculosis.