New derivatives of TOL plasmid pWW0.
Two new segregants, PPW1-1 and PPW161-1, of Pseudomonas putida were isolated from the stock cultures PaW85(pWW0) and PaW85(pWW0-161). Strain PPW1-1 had lost its ability to grow on m-xylene but was able to grow on m-toluate. A deletion of the left-hand of transposon Tn4651, including the upper-operon genes, had taken place in plasmid pWW0mut1, isolated from strain PPW1-1. Additional deletions were observed in pWW0mut1 after 'benzoate-curing' (plasmids pWW0mut15, pWW0mut19, pWW0mut27). The genes of the upper-operon and beginning of the meta-operon were deleted from pWW0-161mut1, isolated from strain PPW161-1. Despite this deletion, cells of PPW161-1 grew on all normal TOL plasmid substrates. The Tol+ phenotype was stable in cells of PPW161-1 growing on benzoate. We propose that this is because in cells of strain PPW161-1 the catabolic genes deleted from pWW0-161mut1 were integrated into the chromosome at the site where the (chromosomally encoded) ortho-pathway genes are located, resulting in the inability of the cells to use this pathway.
Sarand
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I
Institute of Molecular and Cell Biology, University of Tartu, Republic of Estonia.
Mäe
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A
Vilu
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R
Heinaru
A
A
D001565:Benzoates / Q000378:metabolism; D050557:Catechol 2,3-Dioxygenase; D049308:Dioxygenases; D017353:Gene Deletion; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial; D016254:Mutagenesis, Insertional; D009876:Operon; D010105:Oxygenases / Q000032:analysis; D010957:Plasmids; D016958:Pseudomonas putida / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D015183:Restriction Mapping; D014050:Toluene / Q000378:metabolism; D014992:Xylenes / Q000378:metabolism
D001565:Benzoates; D014992:Xylenes; D014050:Toluene; D010105:Oxygenases; D049308:Dioxygenases; D050557:Catechol 2,3-Dioxygenase