Rapid PCR amplification of minimal enediyne polyketide synthase cassettes leads to a predictive familial classification model

Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):11959-63. doi: 10.1073/pnas.2034291100. Epub 2003 Oct 3.

Abstract

A universal PCR method for the rapid amplification of minimal enediyne polyketide synthase (PKS) genes and the application of this methodology to clone remaining prototypical genes from producers of structurally determined enediynes in both family types are presented. A phylogenetic analysis of the new pool of bona fide enediyne PKS genes, consisting of three from 9-membered producers (neocarzinostatin, C1027, and maduropeptin) and three from 10-membered producers (calicheamicin, dynemicin, and esperamicin), reveals a clear genotypic distinction between the two structural families from which to form a predictive model. The results from this study support the postulation that the minimal enediyne PKS helps define the structural divergence of the enediyne core and provides the key tools for generating enediyne hybrid genes/molecular scaffolds; by using the model, a classification is also provided for the unknown enediyne PKS genes previously identified via genome scanning.

Publication types

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

MeSH terms

  • Actinomycetales / enzymology
  • Actinomycetales / genetics
  • Amino Acid Sequence
  • Anti-Bacterial Agents / biosynthesis
  • Anti-Bacterial Agents / chemistry
  • Base Sequence
  • DNA, Bacterial / genetics
  • Escherichia coli / genetics
  • Genes, Bacterial
  • Micromonospora / enzymology
  • Micromonospora / genetics
  • Models, Genetic
  • Molecular Sequence Data
  • Multienzyme Complexes / chemistry
  • Multienzyme Complexes / classification*
  • Multienzyme Complexes / genetics*
  • Phylogeny
  • Polymerase Chain Reaction / methods*
  • Protein Structure, Tertiary
  • Sequence Homology, Amino Acid

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Multienzyme Complexes

Associated data

  • GENBANK/AY162971
  • GENBANK/AY267372
  • GENBANK/AY271660