Genetic analysis of a transposon carrying toluene degrading genes on a TOL plasmid pWWO

Masataka Tsuda, Tetsuo Iino

Research output: Contribution to journalArticlepeer-review

78 Citations (Scopus)

Abstract

Toluene degrading (xyl) genes on a Pseudomonas TOL plasmid pWWO are located within a 39-kb DNA portion. The 56-kb region including these xyl genes and its 17-kb derivative with a deletion of the internal 39-kb portion transposed to various sites on target replicons such as pACYC184 and R388 in escherichia coli recA strains. Thus the 56- and 17-kb regions were designated Tn 4651 and Tn 4652, respectively. Genetic analysis of Tn 4652 demonstrated that its transposition occurs by a two-step process, namely, cointegrate formation and its subsequent resolution. The presence in cis of DNA sequences of no more than 150 bp at both ends of Tn 4652 was prerequisite for cointegrate formation, and this step was mediated by a trans-acting factor, transposase, which was encoded in a 3.0-kb segment at one end of the transposon. Cointegrate resolution took place site-specifically within a 200-bp fragment, which was situated 10 kb away from the transposase gene. Based on the stability of cointegrates formed by various mini-Tn 4652 derivatives, it was shown that the cointergrate resolution requires two trans-acting factors encoded within 1.0- and 1.2-kb fragments that encompass the recombination site involved in the resolution.

Original languageEnglish
Pages (from-to)270-276
Number of pages7
JournalMGG Molecular & General Genetics
Volume210
Issue number2
DOIs
Publication statusPublished - 1987 Dec 1
Externally publishedYes

Keywords

  • TOL plasmid
  • Tn 4651
  • Tn 4652
  • Toluene degrading genes
  • Transposon

ASJC Scopus subject areas

  • Genetics

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