Cloning and expression of the gene for bacteriophage T7 RNA polymerase
Abstract
This application describes a means to clone a functional gene for bacteriophage T7 RNA polymerase. Active T7 RNA polymerase is produced from the cloned gene, and a plasmid has been constructed that can produce the active enzyme in large amounts. T7 RNA polymerase transcribes DNA very efficiently and is highly selective for a relatively long promoter sequence. This enzyme is useful for synthesizing large amounts of RNA in vivo or in vitro, and is capable of producing a single RNA selectively from a complex mixture of DNAs. The procedure used to obtain a clone of the R7 RNA polymerase gene can be applied to other T7-like phages to obtain clones that produce RNA polymerases having different promoter specificities, different bacterial hosts, or other desirable properties. T7 RNA polymerase is also used in a system for selective, high-level synthesis of RNAs and proteins in suitable host cells.
- Inventors:
-
- Stony Brook, NY
- Basel, CH
- Setauket, NY
- Waterloo, CA
- Bellport, NY
- Issue Date:
- Research Org.:
- Associated Universities, Inc., Upton, NY (United States)
- OSTI Identifier:
- 871257
- Patent Number(s):
- 5693489
- Application Number:
- 08/259,560
- Assignee:
- Associated Universities, Inc. (Washington, DC)
- Patent Classifications (CPCs):
-
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
- DOE Contract Number:
- AC02-76CH00016
- Resource Type:
- Patent
- Country of Publication:
- United States
- Language:
- English
- Subject:
- cloning; expression; bacteriophage; t7; rna; polymerase; application; describes; means; clone; functional; active; produced; cloned; plasmid; constructed; produce; enzyme; amounts; transcribes; dna; efficiently; highly; selective; relatively; promoter; sequence; useful; synthesizing; vivo; vitro; capable; producing; single; selectively; complex; mixture; dnas; procedure; obtain; r7; applied; t7-like; phages; clones; polymerases; specificities; bacterial; hosts; desirable; properties; high-level; synthesis; rnas; proteins; suitable; host; cells; bacteriophage t7; bacterial host; application describes; suitable host; rna polymerase; host cells; t7 rna; highly selective; rna polymerases; desirable properties; host cell; promoter specificities; promoter sequence; rna selectively; t7-like phages; single rna; active t7; bacterial hosts; produce rna; polymerase transcribes; active enzyme; obtain clones; complex mixture; transcribes dna; high-level synthesis; rna poly; /435/
Citation Formats
Studier, F William, Davanloo, Parichehre, Rosenberg, Alan H, Moffatt, Barbara A, and Dunn, John J. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. United States: N. p., 1997.
Web.
Studier, F William, Davanloo, Parichehre, Rosenberg, Alan H, Moffatt, Barbara A, & Dunn, John J. Cloning and expression of the gene for bacteriophage T7 RNA polymerase. United States.
Studier, F William, Davanloo, Parichehre, Rosenberg, Alan H, Moffatt, Barbara A, and Dunn, John J. Tue .
"Cloning and expression of the gene for bacteriophage T7 RNA polymerase". United States. https://www.osti.gov/servlets/purl/871257.
@article{osti_871257,
title = {Cloning and expression of the gene for bacteriophage T7 RNA polymerase},
author = {Studier, F William and Davanloo, Parichehre and Rosenberg, Alan H and Moffatt, Barbara A and Dunn, John J},
abstractNote = {This application describes a means to clone a functional gene for bacteriophage T7 RNA polymerase. Active T7 RNA polymerase is produced from the cloned gene, and a plasmid has been constructed that can produce the active enzyme in large amounts. T7 RNA polymerase transcribes DNA very efficiently and is highly selective for a relatively long promoter sequence. This enzyme is useful for synthesizing large amounts of RNA in vivo or in vitro, and is capable of producing a single RNA selectively from a complex mixture of DNAs. The procedure used to obtain a clone of the R7 RNA polymerase gene can be applied to other T7-like phages to obtain clones that produce RNA polymerases having different promoter specificities, different bacterial hosts, or other desirable properties. T7 RNA polymerase is also used in a system for selective, high-level synthesis of RNAs and proteins in suitable host cells.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Dec 02 00:00:00 EST 1997},
month = {Tue Dec 02 00:00:00 EST 1997}
}
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.
journal, February 1985
- Tabor, S.; Richardson, C. C.
- Proceedings of the National Academy of Sciences, Vol. 82, Issue 4
Organization and Expression of Bacteriophage T7 DNA
journal, January 1983
- Studier, F. W.; Dunn, J. J.
- Cold Spring Harbor Symposia on Quantitative Biology, Vol. 47, Issue 0
Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements
journal, June 1983
- Dunn, John J.; Studier, F. William; Gottesman, M.
- Journal of Molecular Biology, Vol. 166, Issue 4, p. 477-535
T7 RNA polymerase directed expression of the Escherichia coli rrnB operon.
journal, May 1986
- Steen, R.; Dahlberg, A. E.; Lade, B. N.
- The EMBO Journal, Vol. 5, Issue 5
Nucleotide sequence of the gene for bacteriophage T7 RNA polymerase
journal, February 1984
- Moffatt, Barbara A.; Dunn, John J.; Studier, F. William
- Journal of Molecular Biology, Vol. 173, Issue 2
A bacteriophage lambda vector for cloning with BamHI and Sau3A
journal, December 1982
- Mizusawa, Saeko; Ward, Douglas F.
- Gene, Vol. 20, Issue 3