Document Detail


Efficient expression of histidine-tagged large hepatitis delta antigen in baculovirus-transduced baby hamster kidney cells.
MedLine Citation:
PMID:  16570347     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
AIM: To study the baculovirus/mammalian cell system for efficient expression of functional large hepatitis delta antigen (L-HDAg).
METHODS: A recombinant baculovirus expressing histidine-tagged L-HDAg (L-HDAgH) was constructed to transduce baby hamster kidney (BHK) cells by a simplified transduction protocol.
RESULTS: The recombinant baculovirus transduced BHK cells with efficiencies higher than 90% as determined by flow cytometry. The expression level was significantly higher than that obtained by plasmid transfection and was further enhanced 3-fold to around 19 pg/cell by the addition of 10 mmol/L sodium butyrate. Importantly, the expressed L-HDAgH was localized to the cell nucleus and correctly isoprenylated as determined by immunofluorescence labeling and confocal microscopy. Moreover, L-HDAgH interacted with hepatitis B surface antigen to form virus-like particles.
CONCLUSION: The fusion with histidine tags as well as overexpression of L-HDAgH in the baculovirus-transduced BHK cells does not impair the biological functions. Taken together, the baculovirus/mammalian cell system offers an attractive alternative for high level expression of L-HDAgH or other proteins that require extensive post-translational modifications.
Authors:
Ying-Wei Chiang; Jaw-Chin Wu; Kuei-Chun Wang; Chia-Wei Lai; Yao-Chi Chung; Yu-Chen Hu
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  World journal of gastroenterology : WJG     Volume:  12     ISSN:  1007-9327     ISO Abbreviation:  World J. Gastroenterol.     Publication Date:  2006 Mar 
Date Detail:
Created Date:  2006-03-29     Completed Date:  2006-08-22     Revised Date:  2014-08-15    
Medline Journal Info:
Nlm Unique ID:  100883448     Medline TA:  World J Gastroenterol     Country:  China    
Other Details:
Languages:  eng     Pagination:  1551-7     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Baculoviridae / genetics*
Cells, Cultured
Cricetinae
DNA, Recombinant
DNA, Viral / analysis
Hepatitis Viruses / genetics,  immunology
Hepatitis delta Antigens / genetics*
Histidine
Kidney
Plasmids
Protein Processing, Post-Translational*
Recombinant Proteins
Transduction, Genetic*
Chemical
Reg. No./Substance:
0/DNA, Recombinant; 0/DNA, Viral; 0/Hepatitis delta Antigens; 0/Recombinant Proteins; 0/hepatitis delta virus large antigen; 4QD397987E/Histidine

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


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