Purification and properties of poliovirus RNA polymerase expressed in Escherichia coli
A cDNA clone encoding the RNA polymerase of poliovirus has been expressed in Escherichia coli under the transcriptional control of a T7 bacteriophage promoter. This poliovirus enzyme was designed to contain only a single additional amino acid, the N-terminal methionine. The recombinant enzyme has been purified to near homogeneity, and polyclonal antibodies have been prepared against it. The enzyme exhibits poly(A)-dependent oligo(U)-primed ply(U) polymerase activity as well as RNA polymerase activity. In the presence of an oligo(U) primer, the enzyme catalyzes the synthesis of a full-length copy of either poliovirus or globin RNA templates. In the absence of added primer, RNA products up to twice the length of the template are synthesized. When incubated in the presence of a single nucleoside triphosphate, (..cap alpha..-/sup 32/P)UTP, the enzyme catalyzes the incorporation of radioactive label into template RNA. These results are discussed in light of previously proposed models of poliovirus RNA synthesis in vitro.
- Research Organization:
- American Cyanamid Co., Pearl River, NY (USA)
- OSTI ID:
- 6204509
- Journal Information:
- J. Virol.; (United States), Journal Name: J. Virol.; (United States) Vol. 63:1; ISSN JOVIA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
BACTERIA
BACTERIOPHAGES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
CARBOXYLIC ACIDS
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DNA
DRUGS
ENZYME ACTIVITY
ENZYMES
ESCHERICHIA COLI
GENE REPRESSORS
ISOTOPES
LIGHT NUCLEI
LIPOTROPIC FACTORS
METHIONINE
MICROORGANISMS
MOLECULAR STRUCTURE
NUCLEI
NUCLEIC ACIDS
NUCLEOPROTEINS
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLIO VIRUS
POLYMERASES
PROTEINS
PURIFICATION
RADIOISOTOPES
RECOMBINANT DNA
RNA POLYMERASES
TRANSFERASES
VIRUSES