Characterization of the cryptic plasmid pSBO2 isolated from Streptococcus bovis JB1 and construction of a new shuttle vector.
A cryptic plasmid designated pSBO2 (3582 bp) was isolated from Streptococcus bovis JB1. The pSBO2 contained putative sites for a double-strand origin (dso), a small transcriptional repressor protein (Cop), countertranscribed RNAs (ctRNAs), and a replication protein (Rep), which were similar to those from pMV158 and pLS1, which were isolated from S. agalactiae, and pWVO1, isolated from Lactococcus lactis. The putative single-strand origin (sso) of pSBO2 was similar to pER341 and pST1, which were isolated from S. thermophilus. Recombinant plasmid designated pSBE2 was constructed to bind pECM184 vector and the DNA fragment containing sso, dso, Cop, ctRNAs, and Rep of pSBO2. When pSBE2 was introduced into S. bovis 12-U-1 and no8, the plasmids in the transformants had deleted the 160-bp fragment between sso and dso. This plasmid, designated pSBE2A, was capable of transforming Escherichia coli and S. bovis strains 12-U-1 and no8 on high frequency; therefore, pSBE2A is an effective shuttle vector.
Rumen Microbiology Research Team, STAFF-Institute, Tsukuba, Ibaraki, Japan.
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M
Nakamura
K
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Ogata
T
T
Nagamine
K
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Tajima
H
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Matsui
Y
Y
Benno
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000235:genetics; D004261:DNA Replication; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics* / Q000302:isolation & purification; D018741:Replication Origin; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D016985:Streptococcus bovis / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D015534:Trans-Activators; C065826:cop protein, Bacteria; C045264:replication initiator protein; D004265:DNA Helicases