摘要
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 purify Mycobacterium tuberculosis protein Rv3425 solubly expressed in the prokaryotic system,and to preliminarily analyze its antigenicity and specificity by Western-blot technique.Methods Rv3425 gene was obtained by PCR and cloned into pET-DsbC vector.After soluble expression in Escherichia coli and affinity purification,the antigenicity and specificity of DsbC-Rv3425 fusion protein were analyzed by Western-blot.Results Soluble expression of Rv3425 gene in Escherichia coli was obtained.Purified DsbC-Rv3425 fusion protein could react strongly with the serum samples of tuberculosis patients,while no reaction was detected in the healthy controls.Conclusions The results suggest that the DsbC-Rv3425 fusion protein is expressed solubly in Escherichia coli.The recombinant DsbC-Rv3425 antigen has good antigenicity and specificity,and therefore has potential application values in the diagnosis of tuberculosis.
出处
《实用预防医学》
CAS
2014年第12期1409-1411,共3页
Practical Preventive Medicine
基金
国家传染病重大项目(2013ZX-10003-006)
