Use of Tn5tac1 to clone a pel gene encoding a highly alkaline, asparagine-rich pectate lyase isozyme from an Erwinia chrysanthemi EC16 mutant with deletions affecting the major pectate lyase isozymes

J Bacteriol. 1995 Aug;177(15):4553-6. doi: 10.1128/jb.177.15.4553-4556.1995.

Abstract

Erwinia chrysanthemi mutant CUCPB5047, delta(pelA pelE) delta(pelB pelC)::28bp delta(pelX) delta 4bp pehX::omega Cmr, was constructed, mutated with Tn5tac1, and screened for isopropyl-beta-D-thiogalactopyranoside-dependent pectate lyase (Pel) production. A Kmr SacI fragment from the hyperexpressing Pel+ mutant CUCPB5066 was cloned into Escherichia coli and sequenced. The gene identified, pelL, encodes a novel, asparagine-rich, highly alkaline enzyme that is similar in primary structure to PelX and in enzymological properties to PelE.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Dickeya chrysanthemi / enzymology
  • Dickeya chrysanthemi / genetics*
  • Gene Deletion
  • Genes, Bacterial
  • Isoenzymes / genetics*
  • Molecular Sequence Data
  • Mutation*
  • Plasmids
  • Polysaccharide-Lyases / biosynthesis
  • Polysaccharide-Lyases / genetics*

Substances

  • Isoenzymes
  • Polysaccharide-Lyases
  • pectate lyase

Associated data

  • GENBANK/L42248