Article Abstract:
A novel rolling-circle-replicating (RCR) plasmid from Pseudomonas putida P8 has been characterized molecularly and its use as a vector considered. Three cryptic circular plasmids, pPP8-3, pPP8-2, and pPP8-1, were found. A double-strand origin of replication, highly conserved in RCR elements, the first element of this type known in pseudomonads, was identified in pPP8-1, with a putative single-strand origin. A kanamyci-resistance gene was ligated into pPP8-1, and the resulting vector was used successfully for transformation of Escherichia coli and P. putida.
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Article Abstract:
The cti gene of Pseudomonas putida was cloned, sequenced and analyzed. This gene imparts to the organism the ability to isomerize unsaturated fatty acids from the cis to the trans configuration. Expression of the gene in Escherichia coli enables this bacteria to isomerize unsaturated fatty acids. The P8 strain of P. putida has a defective cti gene and is unable to convert cis unsaturated fatty acids to the trans isomer.
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Article Abstract:
The integration of a mini-Tn5 containing a cytochrome c operon has been found to undermine the capacity of Psuedomonas putida p8 to cis/trans isomerize unsaturated fatty acids. Experimental results derived from the mutagenesis of P8 strains revealed that the integration of the c operon disabled the ability of the strain to install heme residue into the cytochrome c-type protein. A close similarity between the cytochorome c operon of Psuedomonas fluorescens and Escherichia coli was also observed.
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