Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
3
|
| pubmed:dateCreated |
1998-6-22
|
| pubmed:abstractText |
We have constructed a novel DNA expression vector based on Semliki Forest virus (SFV). SFV produces non-structural proteins (nsPs) which replicate genomic RNA and amplify the mRNA encoding the structural proteins of SFV. A recombinant cDNA genome of SFV, in which the SFV structural genes were replaced by a polylinker cassette to allow for insertion of heterologous DNA, was placed under the control of a cytomegalovirus immediate-early enhancer/promoter with a polyadenylation signal. Transfection of mammalian cells with this SFV-based plasmid vector, pSFV3-CMV-lacZ-pA, resulted in transient high-level expression of a beta-galactosidase reporter gene. The expression level of beta-galactosidase from pSFV3-CMVlacZ-pA was more than 20-fold higher than that obtained from the plasmid with deleted nsPs genes, pSFV3A5976-lacZ, demonstrating that the nsPs genes were essential for the high level of expression. Substantial beta-galactosidase activity was detected in the medium of pSFV3-CMV-lacZ-pA-transfected cells, suggesting that the overproduction of beta-galactosidase caused cell death and release of the protein into the medium. We have demonstrated a high-level expression of the exogenous beta-galactosidase gene from pSFV3-CMV-lacZ-pA constructed using an SFV replication system.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Mar
|
| pubmed:issn |
0969-7128
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:volume |
5
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
415-8
|
| pubmed:dateRevised |
2006-4-21
|
| pubmed:meshHeading |
pubmed-meshheading:9614563-Animals,
pubmed-meshheading:9614563-Cell Culture Techniques,
pubmed-meshheading:9614563-Cell Death,
pubmed-meshheading:9614563-Cricetinae,
pubmed-meshheading:9614563-DNA, Viral,
pubmed-meshheading:9614563-Gene Amplification,
pubmed-meshheading:9614563-Gene Expression,
pubmed-meshheading:9614563-Genes, Reporter,
pubmed-meshheading:9614563-Genetic Vectors,
pubmed-meshheading:9614563-Semliki forest virus,
pubmed-meshheading:9614563-Viral Nonstructural Proteins,
pubmed-meshheading:9614563-beta-Galactosidase
|
| pubmed:year |
1998
|
| pubmed:articleTitle |
Semliki Forest virus-based DNA expression vector: transient protein production followed by cell death.
|
| pubmed:affiliation |
First Department of Internal Medicine, Nagoya University School of Medicine, Japan.
|
| pubmed:publicationType |
Journal Article
|