Details

Autor(en) / Beteiligte

• Chen, Ying-yu
• Shuang, Bao
• Tan, Ya-xia
• Meng, Min-jie
• Han, Pu
• Mo, Xiao-ning
• Song, Quan-sheng
• Qiu, Xiao-yan
• Luo, Xin
• Gan, Qi-ni
• Zhang, Xin
• Zheng, Ying
• Liu, Shun-ai
• Wang, Xiao-ning
• Zhong, Nan-shan
• Ma, Da-long

Titel

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

Ist Teil von

• Chinese medical journal, 2005-02, Vol.118 (4), p.267-274

Ort / Verlag

China: Center for Human Disease Genomics, Peking University, Beijing 100083, China%Guangzhou Institute of Respiratory Disease, First Affiliated Hospital, Guangzhou Medical College, Guangzhou 510120, China%Institute of Molecular Immunology, First Military Medical University of PLA, Guangzhou 510515, China%Center for Disease Control and Prevention of Guangdong Province, Guangzhou 510300, China%Beijing Ditan Hospital, Beijing 100101, China

Erscheinungsjahr

2005

Quelle

MEDLINE

Beschreibungen/Notizen

• BackgroundThe 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.MethodsThe 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-CoVinfected 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-infeeted Vero E6 cells.It was also detected in the lung tissues from patients with SARS. Treatment with and overexpression of protein X4 inhibited the growth of Balb/c 333 cells as determined by cell counting and MTI""" 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.