Document Detail


Expression of adenovirus type 12 E1b 58-kDa protein in Escherichia coli and production of antibodies raised against a 58-kDa::beta-galactosidase fusion protein.
MedLine Citation:
PMID:  2956155     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
DNA fragments coding for the N-terminal 185 amino acids (aa) and for the entire coding region of the adenovirus (Ad)12 E1b 58-kDa protein have been cloned in a prokaryotic expression vector. The N-terminal region of the 58-kDa viral protein (aa 21-205) is expressed as a beta-galactosidase (beta Gal) fusion protein encoded by plasmid pB58Ngal. Escherichia coli strains transformed with this plasmid synthesize a full-length fusion protein of 150-kDa and two truncated proteins: a 140-kDa protein containing aa 64-205 and a 120-kDa polypeptide containing aa 158-205 of the E1b 58-kDa protein. Antibodies raised against purified fusion proteins specifically immunoprecipitate the E1b 58-kDa protein from Ad12-infected and transformed cells. Bacteria transformed with plasmid pB58 carrying the entire E1b 58-kDa coding region (minus the first N-terminal 20 aa which are replaced by 4 aa of beta Gal) showed dramatically reduced growth properties after induction of 58K gene expression. We have not been able to detect substantial amounts of the 58-kDa protein in these cells. However, the viral 58-kDa polypeptide could be synthesized in vitro from plasmid pB58 in a DNA-dependent translation system from E. coli.
Authors:
K Schughart; B von Wilcken-Bergmann; H Esche
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Gene     Volume:  53     ISSN:  0378-1119     ISO Abbreviation:  Gene     Publication Date:  1987  
Date Detail:
Created Date:  1987-08-28     Completed Date:  1987-08-28     Revised Date:  2006-05-01    
Medline Journal Info:
Nlm Unique ID:  7706761     Medline TA:  Gene     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  173-80     Citation Subset:  IM    
Data Bank Information
Bank Name/Acc. No.:
GENBANK/M16661;  M16662
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MeSH Terms
Descriptor/Qualifier:
Adenovirus Early Proteins
Cell Transformation, Viral
Cells, Cultured
Cloning, Molecular
Escherichia coli / genetics
Gene Expression Regulation
Humans
Molecular Weight
Oncogene Proteins, Viral / genetics*,  immunology,  toxicity
Plasmids
Recombinant Fusion Proteins / immunology*
Recombinant Proteins / immunology*
beta-Galactosidase / genetics,  immunology
Chemical
Reg. No./Substance:
0/Adenovirus Early Proteins; 0/Oncogene Proteins, Viral; 0/Recombinant Fusion Proteins; 0/Recombinant Proteins; EC 3.2.1.23/beta-Galactosidase

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


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