PMID: 1369075Nov 1, 1992

Purification and characterization of a DNA-dependent RNA polymerase from Pseudomonas putida

Bioscience, Biotechnology, and Biochemistry
M Fujita, A Amemura

Abstract

DNA-dependent RNA polymerase (EC 2.7.7.6) was purified from Pseudomonas putida. The enzyme had the typical composition of beta',beta,alpha, and sigma subunits of eubacterial RNA polymerases. The molecular masses of the subunits were 156,000 Da, 151,000 Da, 87,000 Da, and 42,000 Da, respectively, as measured by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The NH2-terminal amino acid residues of the alpha subunit had a marked homology with those of the alpha subunit of Escherichia coli RNA polymerase. The enzyme activity was dependent on ribonucleoside triphosphates, Mg2+, and a DNA template, and was inhibited in vitro by rifampicin. The enzyme activity was maximal in the presence of 10 mM MgCl2. In an in vitro transcription assay using the tac promoter-controlled DNA as a template, the RNA polymerase of P. putida initiated transcription at the same site as that of E. coli.

Citations

Jul 9, 2004·Extremophiles : Life Under Extreme Conditions·Hiroaki KawanoFumiyoshi Abe
Dec 24, 2003·Analytical Biochemistry·Peter KuhlmanAshley Galant
Nov 11, 1999·FEMS Microbiology Letters·H Aramaki, M Fujita
Dec 26, 2001·FEMS Microbiology Letters·H AramakiY Sagara
Nov 11, 2020·Environmental Microbiology·Theetha L PavankumarJohn E Hallsworth

Related Concepts

Biotechnology
Alkalescens-Dispar Group
Hydrogen-Ion Concentration
Early Promoters, Genetic
Protein Conformation
Transcriptase
Gene Expression Regulation, Bacterial
Pseudomonas putida
Homologous Sequences, Amino Acid

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