[1]袁军,陈曜,曹炬,等.肺炎链球菌毒力因子ClpP蛋白酶的体外表达及纯化[J].第三军医大学学报,2008,30(18):1753-1755.
 YUAN Jun,CHEN Yao,CAO Ju,et al.Expression and purification of ClpP of Streptococcus pneumoniae in vitro[J].J Third Mil Med Univ,2008,30(18):1753-1755.
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肺炎链球菌毒力因子ClpP蛋白酶的体外表达及纯化(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
30卷
期数:
2008年第18期
页码:
1753-1755
栏目:
论著
出版日期:
2008-09-24

文章信息/Info

Title:
Expression and purification of ClpP of Streptococcus pneumoniae in vitro
作者:
袁军陈曜曹炬尹一兵
重庆医科大学:医学检验系,临床检验诊断学省部共建教育部重点实验室,附属第二医院感染科
Author(s):
YUAN Jun CHEN Yao CAO Ju YIN Yi-Bing
Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Faculty of Laboratory Medicine,  Chongqing 400016, Department of Infectious Diseases, Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China
关键词:
毒力因子肺炎链球菌ClpP蛋白
Keywords:
virulence factory streptococcus pneumonia ClpP
分类号:
Q503;Q556.9;R378.12
文献标志码:
A
摘要:
目的    构建ClpP原核表达载体,获取原核表达的ClpP蛋白分子。    方法    分离培养TIGR4型肺炎链球菌,获取其染色体DNA,采用基因体外重组的方法将完整的ClpP读码框克隆到pET-32a原核表达载体, 并经测序鉴定,所表达的抗原蛋白用Western blot鉴定,镍柱纯化并透析复性,真空冷冻干燥保存。    结果    测序结果证实克隆的基因与GenBank中的序列相符, Western blot证实获得了ClpP 融合蛋白。    结论    获得了高表达的重组抗原蛋白, 经镍柱纯化后纯度可达90%以上。
Abstract:
Objective    To obtain purified ClpP molecule produced by its prokaryotic expression system.     Methods    Template DNA was isolated from cultured TIGR4 Streptococcus pneumoniae by gene recombination technology in vitro. Complete ClpP open reading frame (ORF) was cloned into pET-32a expression vector after being confirmed by sequencing. Antigen protein expressed was identified by Western blotting, purified through Ni2+ affinity chromatography and processed by dialysis and final vacuum freeze drying preservation.     Results    The DNA sequence analysis confirms that the cloned ClpP gene was identical to that in GenBank. ClpP fusion protein was proved by Western blot analysis.     Conclusion    A highly expressed recombinant ClpP fusion antigen protein is successfully obtained, which establishes a material basis on exploring the function of ClpP protein and the mechanism of preventing infection of  streptococcus pneumoniae by ClpP.

参考文献/References:

袁军,陈曜,曹炬,等.   肺炎链球菌毒力因子ClpP蛋白酶的体外表达及纯化[J]. 第三军医大学学报, 2008, 30(18):1753-1755.

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更新日期/Last Update: 2008-09-11