Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
|
| pubmed:dateCreated |
1989-10-18
|
| pubmed:abstractText |
The capability of an Epstein-Barr virus hybrid vector (EBV-CMV), containing the cytomegalovirus (CMV) immediate early enhancer and simian virus 40 promoter, to produce large amounts of authentic mammalian proteins was studied. The cDNA of influenza virus hemagglutinin (HA), a cell surface glycoprotein, was inserted into this vector and the EBV-CMV-HA plasmid was transfected into two human and two monkey cell lines. Southern-blot analysis revealed that the EBV-CMV-HA plasmid was maintained in extrachromosomal state and the recombinant cell clones contained on the average three copies (range 1-24) of the transfected DNA. The recombinant HA polypeptides from different cell clones, selected either randomly or by fluorescence-activated cell sorter, were analysed using immunological techniques. Three of the four cell lines expressed recombinant HA on the cell surface in glycosylated form. The highest production levels, 11.5 micrograms/10(6) cells, were obtained in HeLa cells containing only two copies of EBV-CMV-HA DNA per cell. The protein levels correlated with the mRNA levels in Northern-blot analysis. A corresponding vector, containing the same regulatory signals for HA expression, but lacking the EBV sequences, yielded clones with significantly lower expression levels. The results confirm that the extrachromosomal EBV-CMV vector is very useful in the production of apparently authentic mammalian glycoproteins.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
May
|
| pubmed:issn |
0378-1119
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
30
|
| pubmed:volume |
78
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
287-96
|
| pubmed:dateRevised |
2006-11-15
|
| pubmed:meshHeading |
pubmed-meshheading:2550325-Animals,
pubmed-meshheading:2550325-Cell Line,
pubmed-meshheading:2550325-Cytomegalovirus,
pubmed-meshheading:2550325-DNA, Viral,
pubmed-meshheading:2550325-Electrophoresis, Polyacrylamide Gel,
pubmed-meshheading:2550325-Fluorescent Antibody Technique,
pubmed-meshheading:2550325-Gene Amplification,
pubmed-meshheading:2550325-Genetic Vectors,
pubmed-meshheading:2550325-Hemagglutinin Glycoproteins, Influenza Virus,
pubmed-meshheading:2550325-Hemagglutinins, Viral,
pubmed-meshheading:2550325-Herpesvirus 4, Human,
pubmed-meshheading:2550325-Humans,
pubmed-meshheading:2550325-Immunoblotting,
pubmed-meshheading:2550325-Immunoenzyme Techniques,
pubmed-meshheading:2550325-Mammals,
pubmed-meshheading:2550325-Orthomyxoviridae,
pubmed-meshheading:2550325-Plasmids,
pubmed-meshheading:2550325-Precipitin Tests,
pubmed-meshheading:2550325-Sequence Homology, Nucleic Acid,
pubmed-meshheading:2550325-Transfection
|
| pubmed:year |
1989
|
| pubmed:articleTitle |
Efficient synthesis of influenza virus hemagglutinin in mammalian cells with an extrachromosomal Epstein-Barr virus vector.
|
| pubmed:affiliation |
Orion Genetic Engineering Laboratory, Orion Corporation Ltd., Helsinki, Finland.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|