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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1-2
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pubmed:dateCreated |
1997-11-19
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pubmed:abstractText |
Use of EBV-based vector systems has been limited by the requirement to generate EBNA+ cells which are 'permissive' for replication of an oriP-vector. In current constructs, selectable marker and EBNA-1 are not always co-expressed. This is a significant problem since the EBNA-1 gene product can be toxic in some cell types and may be selected against. In this paper, we describe a gene construct that overcomes this limitation. We have exploited the piconaviral internal ribosome entry site to allow the genes for Epstein-Barr nuclear antigen-1 and G-418 resistance to be transcribed as a dicistronic fusion mRNA under the control of the phosphoglucokinase promoter. This construct can be routinely integrated into human cell lines. The presence of EBNA-1 protein was reflected by a large increase in transfection frequencies (1000-fold) using an oriP-based vector which was shown to replicate stably in these cells with no apparent gross rearrangements detected after 8 weeks in culture. Using this system, G-418 resistance should directly reflect integration, as well as expression of the EBNA-1 gene, which, in turn, increases transfection frequencies and stability of EBV-based vector systems and should result in its increased use.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0378-1119
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:day |
15
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pubmed:volume |
197
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
83-9
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:9332352-Binding Sites,
pubmed-meshheading:9332352-Carcinoma,
pubmed-meshheading:9332352-DNA Replication,
pubmed-meshheading:9332352-Drug Resistance, Microbial,
pubmed-meshheading:9332352-Encephalomyocarditis virus,
pubmed-meshheading:9332352-Epstein-Barr Virus Nuclear Antigens,
pubmed-meshheading:9332352-Genes,
pubmed-meshheading:9332352-Genetic Vectors,
pubmed-meshheading:9332352-Gentamicins,
pubmed-meshheading:9332352-Humans,
pubmed-meshheading:9332352-Kanamycin Kinase,
pubmed-meshheading:9332352-Lung Neoplasms,
pubmed-meshheading:9332352-Phosphotransferases (Alcohol Group Acceptor),
pubmed-meshheading:9332352-Promoter Regions, Genetic,
pubmed-meshheading:9332352-RNA, Messenger,
pubmed-meshheading:9332352-Replication Origin,
pubmed-meshheading:9332352-Ribosomes,
pubmed-meshheading:9332352-Transcription, Genetic,
pubmed-meshheading:9332352-Transfection,
pubmed-meshheading:9332352-Tumor Cells, Cultured
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pubmed:year |
1997
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pubmed:articleTitle |
Improved EBV-based shuttle vector system: dicistronic mRNA couples the synthesis of the Epstein-Barr nuclear antigen-1 protein to neomycin resistance.
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pubmed:affiliation |
Division of Molecular Biology, Roslin Institute, Edinburgh, Scotland, UK.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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