Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis

丛延广; 刘利; 陈志瑾; 饶贤才; 胡晓梅; 李明; 胡勇; 胡福泉

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Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis

更新时间：2026-01-09

- Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis
- Journal of Army Medical University Vol. 29, Issue 15, Pages: 1473-1476(2007)
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  CLC： R378.911;R392-33;R392.11
- Published：2007
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丛延广,刘利,陈志瑾,饶贤才,胡晓梅,李明,胡勇,胡福泉.结核分枝杆菌PstS1-U1融合抗原的表达、纯化及其免疫活性测定[J].陆军军医大学学报(原第三军医大学学报),2007,29(15):1473-1476.

　CONG Yan-guang,LIU Li,CHEN Zhi-jin,RAO Xian-cai,HU Xiao-mei,LI Ming,HU Yong,HU Fu-quan.Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(15):1473-1476.
丛延广,刘利,陈志瑾,饶贤才,胡晓梅,李明,胡勇,胡福泉.结核分枝杆菌PstS1-U1融合抗原的表达、纯化及其免疫活性测定[J].陆军军医大学学报(原第三军医大学学报),2007,29(15):1473-1476. DOI：

　CONG Yan-guang,LIU Li,CHEN Zhi-jin,RAO Xian-cai,HU Xiao-mei,LI Ming,HU Yong,HU Fu-quan.Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(15):1473-1476. DOI：

摘要

目的为获得一种适于本地结核病临床诊断用抗原，将结核分枝杆菌38×10

抗原——即磷酸盐传送蛋白1（phosphate transport system protein l

PstS1）和尿蛋白1(urine protein 1

U1)进行融合表达、纯化并测定其抗原活性。方法用基因拼接法将结核分枝杆菌编码PstS1和U1蛋白的基因拼接在一起，克隆到原核表达载体pET28a中，通过IPTG诱导实现其高效表达，采用亲和层析纯化表达产物。用纯化的融合抗原免疫家兔获得抗血清，用该血清以及活动性肺结核患者血清对纯化产物的抗原活性进行测定。结果成功构建了原核表达质粒，对表达产物进行亲和层析纯化，获得相对分子质量为57×10

的目的蛋白。经Western blot检测融合抗原能够与活动性肺结核患者血清中的抗体发生结合。结论本研究表达并纯化出PstS1-U1融合抗原，为进一步的应用奠定基础。

Abstract

Objective To express and purify Mycobacterium tuberculosis 38kDa (phosphate transport system protein 1

PstS1)-U1(urine protein 1) fusion antigen and determine its immunological activity. Methods The PstS1-U1 fusion gene was amplified from genomic DNA of Mycobacterium tuberculosis by Gene SOEing methods and cloned into pET28a--a prokaryotic expression plasmid. The recombinant PstS1-U1 fusion protein was expressed and purified by affinity chromatography. The immunological activity of purified fusion protein was determined by Western blot with serum of active tuberculosis patie

nts. Results The pET28a-PstS1-U1 expression plasmid was constructed successfully and the expressed fusion protein was 57×10

in molecular weight. Western blot indicated that the purified protein was able to react with antibody in serum of active tuberculosis patients. Conclusion The PstS1-U1 fusion antigen was expressed and purified

which lays the foundation for its potential application in the serodiagnosis of tuberculosis.

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