Document Detail


Expression and characterization of hepatitis C Virus E2 glycoprotein fused to hepatitis B virus preS1(21-47) fragment in CHO cells.
MedLine Citation:
PMID:  12098759     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
To stably express hepatitis C virus (HCV) E2 glycoprotein in CHO cells and facilitate the detection and purification of the expression products, the gene fragment encoding N-terminal 277 amino acids of this protein was fused to the fragment encoding hepatitis B virus (HBV) preS1(21-47) region and inserted into a secretion vector pSecTagB. CHO cells transfected with the recombinant plasmid carrying fusion gene were selected under growth pressure of Zeocin. Secreted fusion products and its cell-associated counterpart were detected by Western blot using E2 specific or preS1 specific antibodies. Glycans carried by the expression products were analyzed with glycan-type specific glycosidases. Most of the cell-associated E2 were found to be high-mannose-type glycosylated, while the secreted E2 proteins were found to be mostly complex-type glycosylated, suggesting further modification in Golgi apparatus upon secretion. Primary studies showed that the fusion antigen could be specifically bind to and elute from anti-preS1 antibody coupled Sepharose resin, suggesting that large-scale preparation of the fusion antigen is feasible with an immunoaffinity resin. This work will contribute to the further study of immunological properties of HCV E2 glycoprotein and also to the study of recombinant HBV/HCV vaccine.
Authors:
Chun-Lin Wang; Li-Xin Zhu; Jing Liu; Zu-Chuan Zhang; Yuan Wang; Guang-Di Li
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica     Volume:  34     ISSN:  0582-9879     ISO Abbreviation:  Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao     Publication Date:  2002 Jul 
Date Detail:
Created Date:  2002-07-05     Completed Date:  2002-08-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  20730160R     Medline TA:  Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai)     Country:  China    
Other Details:
Languages:  eng     Pagination:  400-4     Citation Subset:  IM    
Affiliation:
State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200231, China. wangyuan@server.shcnc.ac.cn
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MeSH Terms
Descriptor/Qualifier:
Animals
Blotting, Western
CHO Cells
Cloning, Molecular
Cricetinae
Fluorescent Antibody Technique
Gene Expression
Hepatitis B Surface Antigens / chemistry,  genetics,  metabolism*
Peptide Fragments / genetics,  metabolism*
Plasmids / genetics
Protein Precursors / chemistry,  genetics,  metabolism*
Recombinant Fusion Proteins / genetics,  metabolism
Viral Envelope Proteins / genetics,  metabolism*
Chemical
Reg. No./Substance:
0/Hepatitis B Surface Antigens; 0/Peptide Fragments; 0/Protein Precursors; 0/Recombinant Fusion Proteins; 0/Viral Envelope Proteins; 0/presurface protein 1, hepatitis B surface antigen; 157184-61-7/glycoprotein E2, Hepatitis C virus

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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