Document Detail


Baculovirus-directed high level expression of the hepatitis delta antigen in Spodoptera frugiperda cells.
MedLine Citation:
PMID:  2016595     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The hepatitis delta antigen (HDAg) is a multifunctional protein. It forms the core-like structure of the hepatitis delta virus (HDV) but also enhances replication of HDV in the nucleus of the hepatocyte. A cDNA fragment encoding HDAg was inserted adjacent to the polyhedrin promoter of Autographa californica nuclear polyhedrosis virus present in the baculovirus transfer vector pVL941. After transfection of Spodoptera frugiperda (Sf9) cells a recombinant baculovirus Ac delta 1 was isolated and purified using filter hybridization techniques. Sf9 cells infected with Ac delta 1 express the HDAg as a non-fused, non-glycosylated protein with an abundance of up to 25% of the total cellular protein mass. Immunoblot analysis using a human polyclonal anti-HD conjugate identified a 22K and a 24K protein in the nucleus of Ac delta 1-infected Sf9 cells. Electron microscopic studies using immunogold labelling showed that the recombinant HDAg (recHDAg) was associated with the hetero-chromatin of the Sf9 cells. The recHDAg produced by Sf9 cells elicited anti-HD antibodies in chimpanzees when injected intramuscularly.
Authors:
T Kos; A Molijn; B Blauw; H Schellekens
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of general virology     Volume:  72 ( Pt 4)     ISSN:  0022-1317     ISO Abbreviation:  J. Gen. Virol.     Publication Date:  1991 Apr 
Date Detail:
Created Date:  1991-05-23     Completed Date:  1991-05-23     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0077340     Medline TA:  J Gen Virol     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  833-42     Citation Subset:  IM    
Affiliation:
Department of Chronic and Infectious Diseases, Primate Center, ITRI-TNO, Rijswijk, The Netherlands.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, Viral / genetics*,  immunology,  isolation & purification
Baculoviridae / genetics*
Base Sequence
Blotting, Northern
Blotting, Southern
Blotting, Western
Cell Line
DNA, Viral / genetics,  isolation & purification
Genes, Viral*
Genetic Vectors
Hepatitis Delta Virus / genetics*,  immunology,  ultrastructure
Hepatitis delta Antigens
Microscopy, Immunoelectron
Molecular Sequence Data
Moths
Pan troglodytes
RNA, Viral / genetics,  isolation & purification
Recombinant Proteins / isolation & purification
Restriction Mapping
Transfection
Chemical
Reg. No./Substance:
0/Antigens, Viral; 0/DNA, Viral; 0/Hepatitis delta Antigens; 0/RNA, Viral; 0/Recombinant Proteins; 0/hepatitis delta virus large antigen

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


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