Cloning and expression of the gene for bacteriophage T7 RNA poly

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

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4351723, 4353201, 43525233, C12N 1567, C12N 1500, C12N 1570, C12N 121

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active

056934890

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.

REFERENCES:
Steen et al. (1986) EMBO J. vol. 5(5) pp. 1099-1105.
Tabor et al. (1985) PNAS vol. 82 pp. 1074-1078.
Mizusawa et al. (1982) Gene vol. 20 pp. 317-322.
Davauloo et al. (1984) PNAS vol. 81 pp. 2035-2039.
Mofflett et al., J. Mol. Biol., 173:265-269 (1984).
Studier et al., Dunn et al., J. Mol. Biol., 166: 477-535 (1983).
Studier et al., Cold Spring Harbor Symp. Quant. Biol., 47:999-1007 (1983).

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