Cloning and characterization of a region responsible for the maintenance of megaplasmid pTAV3 of Paracoccus versutus UW1.
Using cointegrate formation, we constructed a basic replicon of the megaplasmid/mini-chromosome pTAV3 of Paracoccus versutus UW1. It is composed of two adjacent modules, responsible for plasmid replication (rep) and partitioning (par). Functional analysis of the par region identified a determinant of incompatibility (inc2), whose presence is crucial for proper partitioning (the partitioning site). Database searches revealed that the only known replicon with significant homology to that of pTAV3 is encoded by the chromosome cII of Rhodobacter sphaeroides 2.4.1. Incompatibility studies showed that closely related basic replicons are also encoded by megaplasmids (above 400 kb) harbored by four strains of P. pantotrophus. Basic replicons of the pTAV3-type are able to maintain large bacterial genomes, therefore they appear to be good candidates for the construction of vectors specific for Alphaproteobacteria.
Dolowy
Patrycja
P
Department of Bacterial Genetics, Institute of Microbiology, Warsaw University, Miecznikowa 1, 02-096 Warsaw, Poland.
Mondzelewski
Jakub
J
Zawadzka
Renata
R
Baj
Jadwiga
J
Bartosik
Dariusz
D
D001483:Base Sequence / Q000235:genetics; D002876:Chromosomes, Bacterial; D003001:Cloning, Molecular; D004261:DNA Replication; D004269:DNA, Bacterial; D024202:Electrophoretic Mobility Shift Assay; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D009876:Operon; D010230:Paracoccus / Q000235:genetics* / Q000254:growth & development; D010957:Plasmids / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D012093:Replicon / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014169:Transformation, Bacterial / Q000235:genetics
D004269:DNA, Bacterial