Escherichia coli RecBCD enzyme: inducible overproduction and reconstitution of the ATP-dependent deoxyribonuclease from purified subunits

Paul E Boehmer, Peter T. Emmerson

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The intracellular levels of the Escherichia coli RecBCD proteins have been amplified by fusing the recBCD genes to the strong tac promoter/operator in the expression vector, pKK223-3. The overproduced proteins occur at levels amounting to approx. 10% of total cellular protein. Strains harbouring these overexpression plasmids have been used to purify the RecB, RecC and RecD protein subunits, as well as the RecBCD holoenzyme. The individually purified protein subunits can be used to reconstitute the ATP-dependent DNase activity of the RecBCD enzyme.

Original languageEnglish (US)
Pages (from-to)1-6
Number of pages6
JournalGene
Volume102
Issue number1
DOIs
StatePublished - Jun 15 1991
Externally publishedYes

Fingerprint

Exodeoxyribonuclease V
Deoxyribonucleases
Protein Subunits
Adenosine Triphosphate
Escherichia coli
Holoenzymes
Escherichia coli Proteins
Proteins
Plasmids
Genes

Keywords

  • functional enzyme
  • overexpression
  • purification
  • Recombinant proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Escherichia coli RecBCD enzyme : inducible overproduction and reconstitution of the ATP-dependent deoxyribonuclease from purified subunits. / Boehmer, Paul E; Emmerson, Peter T.

In: Gene, Vol. 102, No. 1, 15.06.1991, p. 1-6.

Research output: Contribution to journalArticle

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