Tags

Type your tag names separated by a space and hit enter

Transient gene expression in mammalian and mosquito cells using a recombinant Semliki Forest virus expressing T7 RNA polymerase.
Appl Microbiol Biotechnol. 1999 Dec; 53(1):51-6.AM

Abstract

In this report, we describe a novel recombinant Semliki Forest virus (SFV) expressing T7 RNA polymerase (T7-RP), which was shown to drive transient expression of the chloramphenicol acetyltransferase (cat) gene in mammalian and mosquito cells after transfection of plasmids carrying the reporter gene under the control of the T7 promoter. To our knowledge, this is the first description of a T7-RP-based expression in mosquito cells. Expression of the cat gene was significantly enhanced in mammalian cells by inserting the sequence of the encephalomyocarditis virus internal ribosome entry site between the T7 promoter and the 5' end of the cat gene. In mosquito cells, the level of chloramphenicol acetyltransferase activity was increased by flanking the cat gene with the Autographa californica baculovirus p10 gene 5' and 3' untranslated regions. SFV expressing T7-RP appears, therefore, to be an alternative to other virus-based gene-expression systems in mammalian cells and a powerful tool for protein expression in mosquito cells.

Authors+Show Affiliations

Unité des Arbovirus et virus des fièvres hémorragiques Institut Pasteur, Paris, France. kohla@pasteur.frNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10645625

Citation

Kohl, A, et al. "Transient Gene Expression in Mammalian and Mosquito Cells Using a Recombinant Semliki Forest Virus Expressing T7 RNA Polymerase." Applied Microbiology and Biotechnology, vol. 53, no. 1, 1999, pp. 51-6.
Kohl A, Billecocq A, Préhaud C, et al. Transient gene expression in mammalian and mosquito cells using a recombinant Semliki Forest virus expressing T7 RNA polymerase. Appl Microbiol Biotechnol. 1999;53(1):51-6.
Kohl, A., Billecocq, A., Préhaud, C., Yadani, F. Z., & Bouloy, M. (1999). Transient gene expression in mammalian and mosquito cells using a recombinant Semliki Forest virus expressing T7 RNA polymerase. Applied Microbiology and Biotechnology, 53(1), 51-6.
Kohl A, et al. Transient Gene Expression in Mammalian and Mosquito Cells Using a Recombinant Semliki Forest Virus Expressing T7 RNA Polymerase. Appl Microbiol Biotechnol. 1999;53(1):51-6. PubMed PMID: 10645625.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Transient gene expression in mammalian and mosquito cells using a recombinant Semliki Forest virus expressing T7 RNA polymerase. AU - Kohl,A, AU - Billecocq,A, AU - Préhaud,C, AU - Yadani,F Z, AU - Bouloy,M, PY - 2000/1/25/pubmed PY - 2000/1/25/medline PY - 2000/1/25/entrez SP - 51 EP - 6 JF - Applied microbiology and biotechnology JO - Appl. Microbiol. Biotechnol. VL - 53 IS - 1 N2 - In this report, we describe a novel recombinant Semliki Forest virus (SFV) expressing T7 RNA polymerase (T7-RP), which was shown to drive transient expression of the chloramphenicol acetyltransferase (cat) gene in mammalian and mosquito cells after transfection of plasmids carrying the reporter gene under the control of the T7 promoter. To our knowledge, this is the first description of a T7-RP-based expression in mosquito cells. Expression of the cat gene was significantly enhanced in mammalian cells by inserting the sequence of the encephalomyocarditis virus internal ribosome entry site between the T7 promoter and the 5' end of the cat gene. In mosquito cells, the level of chloramphenicol acetyltransferase activity was increased by flanking the cat gene with the Autographa californica baculovirus p10 gene 5' and 3' untranslated regions. SFV expressing T7-RP appears, therefore, to be an alternative to other virus-based gene-expression systems in mammalian cells and a powerful tool for protein expression in mosquito cells. SN - 0175-7598 UR - https://www.unboundmedicine.com/medline/citation/10645625/Transient_gene_expression_in_mammalian_and_mosquito_cells_using_a_recombinant_Semliki_Forest_virus_expressing_T7_RNA_polymerase_ L2 - https://dx.doi.org/10.1007/s002530051613 DB - PRIME DP - Unbound Medicine ER -