The genome structure of Pseudomonas putida: high-resolution mapping and microarray analysis

Environ Microbiol. 2002 Dec;4(12):819-23. doi: 10.1046/j.1462-2920.2002.00313.x.

Abstract

As part of a collaborative project aimed at sequencing and functionally analysing the entire genome of Pseudomonas putida strain KT2440, a physical clone map was produced as an initial resource. To this end, a high-coverage cosmid library was arrayed and ordered by clone hybridizations. Restriction fragments generated by rare-cutting enzymes and plasmids containing the rrn operon and 23S rDNA of Pseudomonas aeruginosa were used as probes and, parts of the cosmids were end-sequenced. This provided the information necessary for merging and comparing the macro-restriction map, cosmid clone order and sequence information, thereby assuring co-linearity of the eventual sequence assembly with the actual genome. A tiling path of clones was selected, from the shotgun clones used for sequencing, for the production of DNA microarrays that represent the entire genome including its non-coding portions.

Publication types

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

MeSH terms

  • Cloning, Molecular
  • Contig Mapping
  • Cosmids / genetics*
  • DNA, Bacterial / genetics*
  • Gene Library*
  • Genome, Bacterial*
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis
  • Pseudomonas putida / genetics*
  • rRNA Operon / genetics

Substances

  • DNA, Bacterial