|本期目录/Table of Contents|

[1]朱佳佳,刘宏鹏,张爱君,等.结核分枝杆菌ESAT-6诊断抗原基因的克隆及同源性分析[J].宁夏医科大学学报,2010,(03):339-342.
 ZHU Jia-jia,LIU Hong-peng,ZHANG Ai-jun,et al.Cloning and Homologous Analysis of the ESAT-6 Gene from Mycobacterium Tuberculosis [J].Ningxia Medical University,2010,(03):339-342.
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结核分枝杆菌ESAT-6诊断抗原基因的克隆及同源性分析(PDF)
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《宁夏医科大学学报》[ISSN:1005-8486/CN:64-1029/R]

卷:
期数:
2010年03期
页码:
339-342
栏目:
论著
出版日期:
2011-12-30

文章信息/Info

Title:
Cloning and Homologous Analysis of the ESAT-6 Gene from Mycobacterium Tuberculosis
作者:
朱佳佳 刘宏鹏 张爱君 丁淑琴
宁夏医科大学科学技术研究中心,银川 750004
Author(s):
ZHU Jia-jia LIU Hong-pengZHANG Ai-jun DING Shu-qin
Medical Testing Department of Ningxia Medical University,Yinchuan 750004
关键词:
结核分枝杆菌 ESAT-6基因 克隆 序列分析
Keywords:
mycobacterium tuberculosis ESAT-6 gene cloning sequencing
分类号:
R378.91+1
DOI:
-
文献标志码:
A
摘要:
目的 获得结核分枝杆菌ESAT-6抗原基因,进行DNA测序、同源性分析,为筛选结核病候选诊断抗原基因奠定基础。 方法 以结核菌标准菌株H37Rv为模板,通过PCR技术扩增出结核分枝杆菌ESAT-6抗原基因,将其重组到pGEM-T载体后进行序列测定和生物信息学分析。 结果 成功扩增出结核分枝杆菌ESAT-6抗原基因,测序表明该片段开放阅读框由288bp组成,与已发表基因核苷酸序列相比,同源性为99%,推导编码氨基酸序列同源性为90%。 结论 成功克隆结核分枝杆菌ESAT-6基因,测序结果表明该片段阅读框完整。
Abstract:
Objective To obtain and analyze characters and homologies of ESAT-6 gene sequence, and lay bases for screening candidate antigen of Mycobacterium tuberculosis. Methods Total RNA was extracted from protoscoles of cysts. The ESAT-6 gene of Mycobacterium tuberculosis was amplified by PCR and recombined into pGEM-T vector for sequencing and analyzing. Results A DNA sequence with an open reading frame of 288bp had been amplified successfully by PCR. Compared with the DNA sequence published, the homologies were 99%, which deduced that the amino acid sequence of ESAT-6 of Mycobacterium tuberculosis were 90% identities. Conclusion ESAT-6 gene was cloned successfully. By analyzing the DNA sequence, we can make sure that the segment of amplification is ESAT-6, and its reading frame is integrity.

参考文献/References:

[1] Katoch VM. Newerdiagnostic techniques for tuberculosis[J]. Indian J Med Res, 2004, 120(4): 418-428.
[2] AL Sorensen, S Nagai, G Houen, et al. Purification and characterization of a low-molecular-mass T-cell antigen secreted by Mycobacterium tuberculosis [J].Infect Immun,1995, 63: 1710-1717.
[3] Brondt L, Oettinger T, Holm A, et al. Key epitopes on the ESAT-6 antigen recognized in mice during the recall of protective immunity to Mycobacterium tuberculosis [J]. J Immunol, 1996, 157: 3527-3533.
[4] Martin JE, Oettinger T, Andersen P. Delayed-Type Hypersensitivity Responses to ESAT-6 and MPT64 from Mycobacterium tuberculosis in the Guinea Pig [J]. Infect Immun, 1998, 66: 3454-3456.
[5] Pathan AA, Wilkinson KA, Klenerman P, et al. Direct Ex Vivo Analysis of Antigen-Specific IFN-Secreting CD4 T Cells in Mycobacterium tuberculosis-Infected Individuals: Associations with Clinical Disease State and Effect of Treatment[J]. J Immunol, 2001, 167:5217-5225.
[6] LalvaniA, Nagvenkar P, Udwadia Z, et al. Enumeration of T cells specific for RD1-encoded antigens suggests a high prevalence of latent Mycobacterium tuberculosis infection in healthy urban Indians[J].J Infect Dis, 2001, 183: 469-477.
[7] Lalvani A, Pathan A, Mcshane H, et al. Pulmonary function tuberculosis and pulmonary vascular disease: Rapid Detection of Mycobacterium tuberculosis Infection by Enumeration of Antigen-specific T Cells[J]. Am J Respir Crit Care Med, 2001, 163: 824-828.

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备注/Memo

备注/Memo:
收稿日期:2009-09-14 基金项目:宁夏医科大学校级项目(项目编号:XM200825) 作者简介:朱佳佳(1981-),男, 安徽人, 助理实验师。 通信作者:丁淑琴(1974-),女(回族),宁夏人,讲师,硕士,从事病原生物检验教学工作。E-mail:md-dshn@163.com
更新日期/Last Update: 2010-03-20