Strategy for cloning large gene assemblages as illustrated using the phenylacetate and polyhydroxyalkanoate gene clusters

Appl Environ Microbiol. 2004 Aug;70(8):5019-25. doi: 10.1128/AEM.70.8.5019-5025.2004.

Abstract

We report an easy procedure for isolating chromosome-clustered genes. By following this methodology, the entire set of genes belonging to the phenylacetic acid (PhAc; 18-kb) pathway as well as those required for the synthesis and mobilization of different polyhydroxyalkanoates (PHAs; 6.4 kb) in Pseudomonas putida U were recovered directly from the bacterial chromosome and cloned into a plasmid for the first time. The transformation of different bacteria with these genetic constructions conferred on them the ability to either degrade PhAc or synthesize bioplastics (PHAs).

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Biotechnology / methods
  • Cloning, Molecular / methods*
  • Multigene Family / genetics*
  • Phenylacetates / metabolism*
  • Plasmids
  • Polyesters / metabolism*
  • Pseudomonas putida / genetics*
  • Pseudomonas putida / metabolism
  • Transformation, Bacterial

Substances

  • Bacterial Proteins
  • Phenylacetates
  • Polyesters
  • phenylacetic acid