[1]丛延广,刘利,陈志瑾,等.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].第三军医大学学报,2007,29(15):1473-1476.
 CONG Yan-guang,LIU Li,CHEN Zhi-jin,et al.Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis[J].J Third Mil Med Univ,2007,29(15):1473-1476.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第15期
页码:
1473-1476
栏目:
论著
出版日期:
2007-08-15

文章信息/Info

Title:
Prokaryotic expression, purification and immunological activity determination of PstS1-U1 fused antigen of Mycobacterium tuberculosis
作者:
丛延广刘利陈志瑾饶贤才胡晓梅李明胡勇胡福泉
第三军医大学基础医学部微生物学教研室,重庆市微生物工程实验室;解放军324医院检验科
Author(s):
CONG Yan-guang LIU Li CHEN Zhi-jin RAO Xian-cai HU Xiao-mei LI Ming HU Yong HU Fu-quan
Department of Microbiology, College of Medicine, Third Military Medical University, Chongqing 400038;Department of Clinical Laboratory, No. 324 Hospital of PLA, Chongqing 400020, China
关键词:
结核分枝杆菌融合抗原原核表达
Keywords:
Mycobacterium tuberculosis fusion antigen prokaryotic expression
分类号:
R378.911; R392-33; R392.11
文献标志码:
A
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38×103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57×103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 patients.     Results    The pET28a-PstS1-U1 expression plasmid was constructed successfully and the expressed fusion protein was 57×103 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.

参考文献/References:

丛延广,刘利,陈志瑾,等. 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]. 第三军医大学学报, 2007, 29(15):1473-1476.

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更新日期/Last Update: 2008-09-26