学科主题基础医学
The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line
Chen, YY; Shuang, B; Tan, YX; Meng, MJ; Han, P; Mo, XN; Song, QS; Qiu, XY; Luo, X; Gan, QN; Zhang, X; Zheng, Y; Liu, SA; Wang, XN; Zhong, NS; Ma, DL
关键词severe acute respiratory syndrome coronavirus protein X4
刊名CHINESE MEDICAL JOURNAL
2005-02-20
118期:4页:267-274
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Medicine, General & Internal
研究领域[WOS]General & Internal Medicine
关键词[WOS]SARS-CORONAVIRUS ; IDENTIFICATION ; DIAGNOSIS ; PATHOLOGY ; UNIQUE ; GENOME ; ASSAY
英文摘要

Background The genome of the severe acute respiratory syndrome-associated coronavirus ( SARS-CoV) includes sequences encoding the putative protein X4 ( ORF8, ORF7a), consisting of 122 amino acids. The deduced sequence contains a probable cleaved signal peptide sequence and a C-terminal transmembrane helix, indicating that protein X4 is likely to be a type I membrane protein. This study was conducted to demonstrate whether the protein X4 was expressed and its essential function in the process of SARS-CoV infection.

Methods The prokaryotic and eukaryotic protein X4-expressing plasmids were constructed. Recombinant soluble protein X4 was purified from E. coli using ion exchange chromatography, and the preparation was injected into chicken for rising specific polyclonal antibodies. The expression of protein X4 in SARS-CoV infected Vero E6 cells and lung tissues from patients with SARS was performed using immunofluorescence assay and immunohistochemistry technique. The preliminary function of protein X4 was evaluated by treatment with and over-expression of protein X4 in cell lines. Western blot was employed to evaluate the expression of protein X4 in SARS-CoV particles.

Results We expressed and purified soluble recombinant protein X4 from E. coli, and generated specific antibodies against protein X4. Western blot proved that the protein X4 was not assembled in the SARS-CoV particles. Indirect immunofluorescence assays revealed that the expression of protein X4 was detected at 8 hours after infection in SARS-CoV-infected Vero E6 cells. It was also detected in the tung tissues from patients with SARS. Treatment with and overexpression of protein X4 inhibited the growth of Balb/c 313 cells as determined by cell counting and MTT assays.

Conclusion The results provide the evidence of protein X4 expression following SARS-CoV infection, and may facilitate further investigation of the immunopathological mechanism of SARS.

语种英语
WOS记录号WOS:000227511600001
引用统计
被引频次:15[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
版本出版稿
条目标识符http://ir.bjmu.edu.cn/handle/400002259/57452
专题北京大学基础医学院_北京大学人类疾病基因研究中心
作者单位1.Peking Univ, Ctr Human Dis Genom, Beijing 100083, Peoples R China
2.Guangzhou Med Coll, Affiliated Hosp 1, Guangzhou Inst Resp Dis, Guangzhou 510120, Peoples R China
3.First Mil Med Univ PLA, Inst Mol Immunol, Guangzhou 510515, Peoples R China
4.Ctr Dis Control & Prevent Guangdong Prov, Guangzhou 510300, Peoples R China
5.Beijing Ditan Hosp, Beijing 100101, Peoples R China
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GB/T 7714
Chen, YY,Shuang, B,Tan, YX,et al. The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line[J]. CHINESE MEDICAL JOURNAL,2005,118(4):267-274.
APA Chen, YY.,Shuang, B.,Tan, YX.,Meng, MJ.,Han, P.,...&Ma, DL.(2005).The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line.CHINESE MEDICAL JOURNAL,118(4),267-274.
MLA Chen, YY,et al."The protein X4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of Balb/c 3T3 cell line".CHINESE MEDICAL JOURNAL 118.4(2005):267-274.
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