| Efficient expression of histidine-tagged large hepatitis delta antigen in baculovirus-transduced baby hamster kidney cells. | |
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MedLine Citation:
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PMID: 16570347 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Ying-Wei Chiang; Jaw-Chin Wu; Kuei-Chun Wang; Chia-Wei Lai; Yao-Chi Chung; Yu-Chen Hu |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: World journal of gastroenterology : WJG Volume: 12 ISSN: 1007-9327 ISO Abbreviation: World J. Gastroenterol. Publication Date: 2006 Mar |
Date Detail:
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Created Date: 2006-03-29 Completed Date: 2006-08-22 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 100883448 Medline TA: World J Gastroenterol Country: China |
Other Details:
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Languages: eng Pagination: 1551-7 Citation Subset: IM |
Affiliation:
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Department of Chemical Engineering, National Tsing Hua University, Hsinchu 300, Taiwan, China. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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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:
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0/DNA, Recombinant; 0/DNA, Viral; 0/Hepatitis delta Antigens; 0/Recombinant Proteins; 0/hepatitis delta virus large antigen; 71-00-1/Histidine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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