Isolation and characterization of pHW15, a small cryptic plasmid from Rahnella genomospecies 2.
A small cryptic plasmid designated pHW15 was isolated from Rahnella genomospecies 2 WMR15 and its complete nucleotide sequence was determined. The plasmid contained 3002 bp with a G+C content of 47.4%. The origin of replication was identified by deletion analysis as a region of about 600 bp. This region had an identity of 70% to the replication origin of the ColE1 plasmid at the nucleotide level. Sequence analysis revealed the typical elements: RNA I, RNA II and their corresponding promoters, a sequence allowing hybridisation of RNA II to the DNA and favouring processing by RNaseH, a single-strand initiation determinant (ssi) that allows initiation of lagging-strand synthesis, and a terH sequence required for termination of lagging-strand synthesis. The plasmid contained three expressed open reading frames, one of which showed homology to a ColE1 plasmid-encoded protein. Furthermore, a multimer resolution site was identified by sequence analysis. Its deletion resulted in formation of plasmid multimers during growth leading to an increased plasmid loss rate.
Rozhon
Wilfried M
WM
Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Dr. Bohrgasse 3, A-1030 Vienna, Austria. wilfried.rozhon@gmi.oeaw.ac.at
Petutschnig
Elena K
EK
Jonak
Claudia
C
D001482:Base Composition; D020029:Base Pairing; D001483:Base Sequence; D016000:Cluster Analysis; D017931:DNA Primers; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics* / Q000302:isolation & purification; D020638:Rahnella / Q000235:genetics*; D018741:Replication Origin / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D017931:DNA Primers