Isolation and molecular characterization of pMG160, a mobilizable cryptic plasmid from Rhodobacter blasticus.
A 3.4-kb cryptic plasmid was obtained from a new isolate of Rhodobacter blasticus. This plasmid, designated pMG160, was mobilizable by the conjugative strain Escherichia coli S17.1 into Rhodobacter sphaeroides, Rhodobacter capsulatus, and Rhodopseudomonas palustris. It replicated in the latter strains but not in Rhodospirillum rubrum, Rhodocyclus gelatinosus, or Bradyrhizobium species. Plasmid pMG160 was stably maintained in R. sphaeroides for more than 100 generations in the absence of selection but showed segregational instability in R. palustris. Instability in R. palustris correlated with a decrease in plasmid copy number compared to the copy number in R. sphaeroides. The complete nucleotide sequence of plasmid pMG160 contained three open reading frames (ORFs). The deduced amino acid sequences encoded by ORF1 and ORF2 showed high degrees of homology to the MobS and MobL proteins that are involved in plasmid mobilization of certain plasmids. Based on homology with the Rep protein of several other plasmids, ORF3 encodes a putative rep gene initiator of plasmid replication. The functions of these sequences were demonstrated by deletion mapping, frameshift analysis, and analysis of point mutations. Two 6.1-kb pMG160-based E. coli-R. sphaeroides shuttle cloning vectors were constructed and designated pMG170 and pMG171. These two novel shuttle vectors were segregationally stable in R. sphaeroides growing under nonselective conditions.
Inui
Masayuki
M
Research Institute of Innovative Technology for the Earth, Kizu, Soraku, Kyoto 619-0292, Japan.
Nakata
Kaori
K
Roh
Jung Hyeob
JH
Vertès
Alain A
AA
Yukawa
Hideaki
H
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D004261:DNA Replication; D004926:Escherichia coli / Q000235:genetics; D017353:Gene Deletion; D018628:Gene Dosage; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics* / Q000302:isolation & purification; D016992:Rhodobacter / Q000235:genetics*; D012242:Rhodobacter sphaeroides / Q000235:genetics; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins