Preparation and characterization of large amounts of restriction fragments containing the E. coli lac control elements

Stephen C. Hardies, Robert D. Wells

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Large quantities of pure DNA fragments (789, 203 and 95 bp in length) containing the Escherichia coli lac controlling elements (operator, promoter, CRP binding site) were prepared from appropriate recombinant plasmids. High pressure liquid chromatography on RPC-5 or preparative sucrose gradient centrifugation was used to fractionate the pVH51 vector from the inserts. The fragments had few, if any, nicks or depurinated sites, and the majority of the fragment ends were intact. Absorbance-temperature profiles on the fragments showed multiphasic transitions.

Original languageEnglish (US)
Pages (from-to)1-14
Number of pages14
JournalGene
Volume7
Issue number1
DOIs
StatePublished - Sep 1979

Keywords

  • CRP binding site
  • Promoter
  • RPC-5 column chromatography
  • T curves
  • operator
  • plasmids
  • restriction endonucleases EcoRI, HeaII, HindIII, AluI

ASJC Scopus subject areas

  • Genetics

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