Purification of Rhodobacter sphaeroides RNA polymerase and its sigma factors

Methods Enzymol. 2003;370:54-65. doi: 10.1016/S0076-6879(03)70005-6.

Abstract

This article summarized methods to obtain RNA polymerase and sigma factors that can be used to analyze the in vitro control of gene expression by the facultative phototroph R. sphaeroides. While not a topic of this article, these purified components also allow one to analyze R. sphaeroides promoters that use activators to stimulate transcription. We expect that these approaches will be increasingly useful as investigators continue to dissect the number of unusual signal transduction pathways that control gene expression in this and other related species.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Biochemistry / methods*
  • Chromatography, Affinity
  • DNA-Directed RNA Polymerases / chemistry*
  • DNA-Directed RNA Polymerases / isolation & purification*
  • Electrophoresis, Polyacrylamide Gel
  • Heat-Shock Proteins / chemistry
  • Molecular Sequence Data
  • Plasmids / metabolism
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Rhodobacter sphaeroides / enzymology*
  • Sigma Factor / chemistry*
  • Signal Transduction
  • Sonication
  • Transcription Factors / chemistry
  • Transcription, Genetic

Substances

  • Heat-Shock Proteins
  • Recombinant Proteins
  • Sigma Factor
  • Transcription Factors
  • heat-shock sigma factor 32
  • sporulation-specific sigma factors
  • RNA polymerase sigma 70
  • DNA-Directed RNA Polymerases