Structural analysis of the 6 kb cryptic plasmid pFAJ2600 from Rhodococcus erythropolis NI86/21 and construction of Escherichia coli-Rhodococcus shuttle vectors.
The complete nucleotide sequence of the 5936 bp cryptic plasmid pFAJ2600 from Rhodococcus erythropolis NI86/21 was determined. Based on the characteristics of its putative replication genes, repA and repB, pFAJ2600 was assigned to the family of pAL5000-related small replicons identified in Mycobacterium (pAL5000), Corynebacterium (pXZ10142), Brevibacterium (pRBL1), Bifidobacterium (pMB1) and Neisseria (pJD1). The replication systems of these plasmids show striking similarities to the ones used by the ColE2 family of plasmids from Enterobacteria with respect to both trans-acting factors and ori sequences. Two possible plasmid stabilization systems are encoded on pFAJ2600: a site-specific recombinase (PmrA) related to the Escherichia coli Xer proteins for plasmid multimer resolution and an ATPase (ParA) related to the A-type of proteins in sop/par partitioning systems. The proposed replication termination region of pFAJ2600 has features in common with the Ter loci of Bacillus subtilis. Chimeras composed of a pUC18-Cmr derivative inserted in the parA-repA intergenic region of vector pFAJ2600 produced vectors that could be shuttled between Escherichia coli and several Rhodococcus species (R. erythropolis, R. fascians, R. rhodochrous, R. ruber). The pFAJ2600-based shuttle vector pFAJ2574 was stably maintained in R. erythropolis and R. fascians growing under non-selective conditions.
De Mot
René
R
F. A. Janssens Laboratory of Genetics, Department of Applied Plant Sciences, Catholic University of Leuven, Willem de Croylaan 42, B-3001 Heverlee, Belgium.
Nagy
István
I
F. A. Janssens Laboratory of Genetics, Department of Applied Plant Sciences, Catholic University of Leuven, Willem de Croylaan 42, B-3001 Heverlee, Belgium.
De Schrijver
Adinda
A
F. A. Janssens Laboratory of Genetics, Department of Applied Plant Sciences, Catholic University of Leuven, Willem de Croylaan 42, B-3001 Heverlee, Belgium.
Pattanapipitpaisal
Pranee
P
F. A. Janssens Laboratory of Genetics, Department of Applied Plant Sciences, Catholic University of Leuven, Willem de Croylaan 42, B-3001 Heverlee, Belgium.
Schoofs
Geert
G
F. A. Janssens Laboratory of Genetics, Department of Applied Plant Sciences, Catholic University of Leuven, Willem de Croylaan 42, B-3001 Heverlee, Belgium.
Vanderleyden
Jos
J
F. A. Janssens Laboratory of Genetics, Department of Applied Plant Sciences, Catholic University of Leuven, Willem de Croylaan 42, B-3001 Heverlee, Belgium.
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D002874:Chromosome Mapping; D004254:DNA Nucleotidyltransferases / Q000235:genetics; D004261:DNA Replication / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005822:Genetic Vectors; D019426:Integrases; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D045522:Recombinases; D018741:Replication Origin; D012093:Replicon; D012240:Rhodococcus / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D045522:Recombinases; C081902:pmrA protein, Bacteria; D004254:DNA Nucleotidyltransferases; D019426:Integrases; C115006:integron integrase IntI1