Purification and characterization of the DNA-dependent RNA polymerase from Clostridium acetobutylicum.

Abstract:

:The DNA-dependent RNA polymerase (EC 2.7.7.6) from Clostridium acetobutylicum DSM 1731 has been purified to homogeneity and characterized. The purified enzyme was composed of four subunits and had a molecular mass of 370,000 Da. Western immunoblot analysis with polyclonal antibodies against the sigma 70 subunit of Escherichia coli RNA polymerase identified the 46,000-Da subunit as an immunologically and probably functionally related protein. The other three subunits of 128,000, 117,000, and 42,000 Da are tentatively analogous to the beta, beta', and alpha subunits, respectively, of other eubacterial RNA polymerases. The RNA polymerase activity was completely dependent on Mg2+, nucleoside triphosphates, and a DNA template. The presence of Mg2+ or Mn2+ in buffers used for purification or storage caused irreversible inactivation of the RNA polymerase.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Pich A,Bahl H

doi

10.1128/jb.173.6.2120-2124.1991

subject

Has Abstract

pub_date

1991-03-01 00:00:00

pages

2120-4

issue

6

eissn

0021-9193

issn

1098-5530

journal_volume

173

pub_type

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