cis-trans isomerization of unsaturated fatty acids: cloning and sequencing of the cti gene from Pseudomonas putida P8.
Transposon mutants of Pseudomonas putida P8 were generated by applying a mini-Tn5 mutagenesis system. The mutants obtained were checked for their ability to tolerate increased temperatures and elevated phenol concentrations. Approximately 5,800 transposon mutants were used to generate a pool of 600 temperature-sensitive strains; one of these strains was identified as being damaged in its ability to perform cis-trans isomerization of fatty acids. A gene library of P. putida P8 was constructed and screened by using as a probe sequences immediately adjacent to the mini-Tn5 insertion. A DNA fragment that complemented the mutation was isolated and cloned. The corresponding gene, termed cti, is located close to the methionine synthase locus (metH) in P. putida P8. A cti-carrying fragment integrated into a plasmid also conferred the ability for cis-trans isomerization to Escherichia coli; the cti gene was completely sequenced, and the amino acid sequence was deduced.
Holtwick
R
R
Institut für Mikrobiologie, Westfälische-Wilhelms-Universität Münster, Germany.
Meinhardt
F
F
Keweloh
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016958:Pseudomonas putida / Q000235:genetics*; D013237:Stereoisomerism
D004251:DNA Transposable Elements; D005231:Fatty Acids, Unsaturated