Molecular, genetic, and functional analysis of the basic replicon of pVA380-1, a plasmid of oral streptococcal origin

LeBlanc, Donald J.; Lee, Linda N.; Abu-Al-Jaibat, Angela

doi:10.1016/0147-619x(92)90044-b

Cited by 193 publications

(124 citation statements)

References 35 publications

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“…Bacterial strains and plasmids used included: L. lactis M189 (Duan et al, 1996); L. lactis LM0230, a plasmidfree derivative of L. lactis 02 (Efstathiou and McKay, 1977); L. lactis LM0230 SmR, a streptomycin resistant (SmR) derivative of LM0230; Escherichia coli JM109 (Sambrook et al, 1989); pSA3 (10.2 kb) encoding resistance to erythromycin (EmR) (Dao and Ferretti, 1985); pDL278 (6.6 kb) encoding resistance to spectinomycin (SpecR) (LeBlanc and Lee, 1992); pMU1327 (7.5 kb) encoding EmR and carrying the cat reporter gene encoding chloramphenicol resistance (CmR) (Achen et al, 1986); and pGEM-7Zf(+) (3.0 kb) encoding resistance to ampicillin (APR) (Promega, Madison, WI, U.S.A.). All lactococcal strains were cultured at 30°C in M17 medium (Terzaghi and Sandine, 1975) containing 0.5% (w/v) glucose (M17G) or lactose (M17L).…”

Section: Methodsmentioning

confidence: 99%

Cloning of a gene encoding nisin resistance from Lactococcus lactis subsp. Iactis M189 which is transcribed from an extended -10 promoter.

Liu

Harvey

Dunn

1997

J. Gen. Appl. Microbiol.

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A 56-kb plasmid was identified in Lactococcus lactic subsp. lactis (L. lactic) M189 which encodes resistance to nisin (NisR) following mobilization of the plasmid into L. lactis LM0230. The NisR determinant was localized on a 1.6-kb Hindlll fragment by DNA restriction fragment deletion and sub-cloning. An open reading frame (ORF) of 957 bases was identified by sequence analysis and its transcription start site was mapped by primer extension. The ORF is flanked by two regions which exhibit complete homology to parts of the inverted repeat sequences of IS981 and ISS1T. The promoter for transcription was found to consist of an extended -10 site (TgTGtTATAAT) that lacks a -35 site . Function of the extended -10 promoter was demonstrated by its ability to express the promoterless cat gene from Staphylococcus aureus. Base substitution analysis revealed that the TgTGt extension is essential for promoter efficiency in L. lactis.

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Section: Methodsmentioning

confidence: 99%

Cloning of a gene encoding nisin resistance from Lactococcus lactis subsp. Iactis M189 which is transcribed from an extended -10 promoter.

Liu

Harvey

Dunn

1997

J. Gen. Appl. Microbiol.

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“…pINY1834, and pADi have been described previously (4,7,9). The E. coli-E. faecalis shuttle vectors pWM402 (35), pDL276, and pDL278 (13,22) have been used extensively in our laboratory for cloning pCF10 DNA (4,5,13,19). Construction of pINY6023 was carried out by removing the 7.5-kb SalI-XbaI fragment of pCF10 indicated in Fig.…”

Section: Methodsmentioning

confidence: 99%

Cloning and characterization of a region of the Enterococcus faecalis conjugative plasmid, pCF10, encoding a sex pheromone-binding function

1993

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In order to investigate the mechanism by which peptide sex pheromones induce expression of the conjugation functions of certain Enterococcus faecalis plasmids, a biological assay was developed to measure the ability of cells carrying the conjugative plasmid pCF10 to bind the sex pheromone cCF10. The data indicated that pCF10 endows its host E. faecalis cell with the ability to specifically remove (apparently by irreversible binding) cCF10 activity from culture medium. The pCF10 DNA encoding this ability was localized to a 3.4-kb segment within a region involved in negative control of expression of conjugal transfer functions. This segment also encoded ability to bind the pheromone inhibitor peptide iCF10. DNA sequencing revealed three open reading frames, which have been denoted prgW (pheromone responsive gene W), prgZ, and prgY. The deduced product of prgW resembled regulatory proteins from other bacteria and eucaryotes, with a very high degree of identity within a putative DNA-binding domain. The prgY gene actually extended into an adjacent region of pCF10 and could encode a protein with significant similarity to a protein called TraB, believed to be involved in shutdown of pheromone cAD1 production by cells carrying the pheromone-inducible hemolysin plasmid pAD1, according to F.Y. An and D.B. Clewell (Abstr. Gen. Meet. Am. Soc. Microbiol. 1992, H70, 1992). The prgZ gene product showed significant relatedness to binding proteins encoded by oligopeptide permease (opp) operons in gram-positive and gram-negative bacteria and is highly similar to a pAD1-encoded protein, TraC, which is believed to mediate sex pheromone cAD1 binding (K. Tanimoto, F. Y. An, and D. B. Clewell, submitted for publication). A Tn5 insertion into prgZ abolished cCF10 binding ability.

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“…Analysis of ssDNA production during replication of the FG2 plasmid replicon To investigate the mode of replication, pND611 was examined for the presence of ssDNA intermediates during replication. The plasmid pDL278, which is known to replicate via the RCR mechanism (LeBlanc et al, 1992), was used as a positive control. No ssDNA was detected with pND611 (a of B and D in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…lactis shuttle vector encoding erythromycin resistance (EmR) (Achen et al, 1986); pDL278 (6.6 kb), an E. coliiL. lactis shuttle vector encoding spectinomycin resistance (SpecR) (LeBlanc et al, 1992); pGEM-7Zf(+) (3.0 kb), an E. coil vector encoding ampicillin resistance (APR) (Promega, Madison, WI, U.S.A.); pUC19 (2.7 kb), an E. coil vector encoding APR (Sambrook et al, 1989); pE194 (3.7 kb), a Staphylococcus aureus plasmid encoding EmR (Horinouchi and Weisblum, 1982); pND628 (7.1 kb), a derivative of pGEM-7Zf(+) carrying a 1.6-kb Hindlll fragment encoding nisin resistance (Ni5R) (GenBank accession code: U25181) and a 2.5-kb Sphl/Xbal fragment containing the replication region from pND608 (this study).…”

Section: Methodsmentioning

confidence: 99%

Cloning and sequence analysis of a plasmid replicon from Lactococcus lactis subsp. cremoris FG2.

Liu

Duan

Dunn

1997

J. Gen. Appl. Microbiol.

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A replication region from one of the Lactococcus lactis subsp. cremoris FG2 plasmids was isolated by cloning of a 4.8-kb Xbal fragment into a replication probe vector and transformation into L. lactis LM0230. A 1.8-kb region within this fragment was sequenced and confirmed by PCR subcloning to encode a functional replicon in LM0230. The replicon consists of an open reading frame encoding a putative replication protein (Rep) of 386 amino acids and a non-coding region (on) which features several structural motifs typical of other known replication origins, including a 22-bp iteron sequence tandemly repeated three and a half times, a 10-bp direct repeat and two sets of inverted repeats. The oni region could drive replication of its plasmid when supplied with the replication region in-trans. The lack of detectable single-stranded DNA during replication and the existence of extensive homology with other known lactococcal theta replicons strongly suggest that this region encodes a theta-replicating mechanism.

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Molecular, genetic, and functional analysis of the basic replicon of pVA380-1, a plasmid of oral streptococcal origin

Cited by 193 publications

References 35 publications

Cloning of a gene encoding nisin resistance from Lactococcus lactis subsp. Iactis M189 which is transcribed from an extended -10 promoter.

Cloning of a gene encoding nisin resistance from Lactococcus lactis subsp. Iactis M189 which is transcribed from an extended -10 promoter.

Cloning and characterization of a region of the Enterococcus faecalis conjugative plasmid, pCF10, encoding a sex pheromone-binding function

Cloning and sequence analysis of a plasmid replicon from Lactococcus lactis subsp. cremoris FG2.

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