RepB proteins of the multipartiteRhizobium leguminosarumbv.trifoliigenome discriminate between centromere-likeparSsequences for plasmid segregational stability

2016 ◽  
Vol 102 (3) ◽  
pp. 446-466 ◽  
Author(s):  
Piotr Koper ◽  
Kamil Żebracki ◽  
Małgorzata Marczak ◽  
Anna Skorupska ◽  
Andrzej Mazur
Keyword(s):  
Segregational Stability

scholarly journals Replication of Staphylococcal Multiresistance Plasmids

2000 ◽  
Vol 182 (8) ◽  
pp. 2170-2178 ◽  
Author(s):  
Neville Firth ◽  
Sumalee Apisiridej ◽  
Tracey Berg ◽  
Brendon A. O'Rourke ◽  
Steve Curnock ◽  
...  
Keyword(s):  
Sequence Similarity ◽  
Heavy Metal Resistance ◽  
Metal Resistance ◽  
Replication Initiation ◽  
Conjugative Plasmid ◽  
Rolling Circle ◽  
Segregational Stability ◽  
Structural And Functional Properties ◽  
Resistance Plasmids ◽  
Genes Encoding

ABSTRACT Based on structural and functional properties, three groups of large staphylococcal multiresistance plasmids have been recognized, viz., the pSK1 family, pSK41-like conjugative plasmids, and β-lactamase–heavy-metal resistance plasmids. Here we describe an analysis of the replication functions of a representative of each of these plasmid groups. The replication initiation genes from theStaphylococcus aureus plasmids pSK1, pSK41, and pI9789::Tn552 were found to be related to each other and to the Staphylococcus xylosus plasmid pSX267 and are also related to rep genes of several plasmids from other gram-positive genera. Nucleotide sequence similarity between pSK1 and pI9789::Tn552 extended beyond theirrep genes, encompassing upstream divergently transcribed genes, orf245 and orf256, respectively. Our analyses revealed that genes encoding proteins related to the deducedorf245 product are variously represented, in several types of organization, on plasmids possessing six seemingly evolutionarily distinct types of replication initiation genes and including both theta-mode and rolling-circle replicons. Construction of minireplicons and subsequent functional analysis demonstrated that orf245is required for the segregational stability of the pSK1 replicon. In contrast, no gene equivalent to orf245 is evident on the conjugative plasmid pSK41, and a minireplicon encoding only the pSK41 rep gene was found to exhibit a segregational stability approaching that of the parent plasmid. Significantly, the results described establish that many of the large multiresistance plasmids that have been identified in clinical staphylococci, which were formerly presumed to be unrelated, actually utilize an evolutionarily related theta-mode replication system.

scholarly journals A Single Gene on the Staphylococcal Multiresistance Plasmid pSK1 Encodes a Novel Partitioning System

2003 ◽  
Vol 185 (7) ◽  
pp. 2143-2152 ◽  
Author(s):  
Alice E. Simpson ◽  
Ronald A. Skurray ◽  
Neville Firth
Keyword(s):  
Single Gene ◽  
Replication Initiation ◽  
Plasmid Replication ◽  
Rolling Circle ◽  
Plasmid Segregation ◽  
Segregational Stability ◽  
Protein Encoding ◽  
Single Protein ◽  
Encoding Gene ◽  
Postsegregational Killing

ABSTRACT The orf245 gene is located immediately upstream of, and divergently transcribed from, the replication initiation gene, rep, of the Staphylococcus aureus multiresistance plasmid pSK1, and related genes have been found in association with a range of evolutionarily distinct replication genes on plasmids from various gram-positive genera. orf245 has been shown previously to extend the segregational stability of a pSK1 minireplicon. Here we describe an investigation into the basis of orf245-mediated stabilization. orf245 was not found to influence transcription of pSK1 rep, indicating that it is not directly involved in plasmid replication. This was confirmed by demonstrating that orf245 is able to enhance the segregational stability of heterologous theta- and rolling-circle-replicating replicons, suggesting that it encodes a plasmid maintenance function. Evidence inconsistent with postsegregational killing and multimer resolution mechanisms was obtained; however, the intergenic region upstream of orf245 was found to mediate orf245-dependent incompatibility, as would be expected if it encodes a cis-acting centromere-like site. Taken together, these findings implicate active partitioning as the probable basis of the activity of orf245, which is therefore redesignated par. Since it is unrelated to any gene known to play a role in plasmid segregation, it seems likely that pSK1 par potentially represents the prototype of a novel class of active partitioning systems that are distinguished by their capacity to enhance plasmid segregational stability via a single protein-encoding gene.

scholarly journals Protection Efficacy of Oral Bait Probiotic Vaccine Constitutively Expressing Tetravalent Toxoids against Clostridium perfringens Exotoxins in Livestock (Rabbits)

Vaccines ◽  
2020 ◽  
Vol 8 (1) ◽  
pp. 17 ◽  
Author(s):  
Jing Bai ◽  
Xinyuan Qiao ◽  
Yingying Ma ◽  
Meijing Han ◽  
Shuo Jia ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Vaccination Strategy ◽  
Opportunistic Pathogen ◽  
Oral Immunization ◽  
Segregational Stability ◽  
Protection Rate ◽  
Protection Efficacy ◽  
Etiological Agents ◽  
Enteritis Necroticans ◽  
Trivalent Vaccine

Clostridium perfringens is an opportunistic pathogen. Its main virulence factors are exotoxins, which are the etiological agents of enteritis necroticans and enterotoxemia caused in livestock (cattle, sheep, and rabbits). Here, we demonstrated effective immune protection for rabbits against α, β, and ε exotoxins of C. perfringens provided by an oral tetravalent bait probiotic vaccine delivering α, ε, β1, and β2 toxoids of C. perfringens. Results showed that the recombinant probiotic had good segregational stability and good colonization ability in the rabbit intestinal tract. Oral administration of the probiotic vaccine can effectively elicit significant levels of antigen-specific mucosa sIgA and sera IgG antibodies with exotoxin-neutralizing activity. Additionally, oral immunization with the probiotic vaccine effectively promoted lymphoproliferation and Th1/Th2-associated cytokine production. The protection rate of immunized rabbits with the probiotic vaccine was 80% after challenging rabbits with a combination of C. perfringens (toxinotypes A, C, and D) and exotoxin mixture, which was better than the 60% provided by a commercial inactivated C. perfringens A, C, and D trivalent vaccine. Moreover, obvious histopathological changes were observed in the intestinal tissues of rabbits in the commercial vaccine and PBS groups. The bait probiotic vaccine can provide effective protection against C. perfringens exotoxins, suggesting a promising C. perfringens vaccination strategy.

Improvement in segregational stability of recombinant plasmids by retransformation of E. coli host cells

1993 ◽  
Vol 15 (8) ◽  
Author(s):  
Jorge Lavastida ◽  
Lilia Colon ◽  
Michael Ward ◽  
BenjaminC. Stark
Keyword(s):  
Host Cells ◽  
E Coli ◽  
Recombinant Plasmids ◽  
Segregational Stability

scholarly journals Localization of the Naturally Occurring Plasmid ColE1 at the Cell Pole

2006 ◽  
Vol 189 (5) ◽  
pp. 1946-1953 ◽  
Author(s):  
Shiyin Yao ◽  
Donald R. Helinski ◽  
Aresa Toukdarian
Keyword(s):  
Plasmid Stability ◽  
Time Lapse ◽  
Replication Initiation ◽  
Stability Studies ◽  
Polar Localization ◽  
Replication Initiation Protein ◽  
Cell Pole ◽  
Segregational Stability ◽  
Dynamic Movement ◽  
Naturally Occurring

ABSTRACT The naturally occurring plasmid ColE1 was found to localize as a cluster in one or both of the cell poles of Escherichia coli. In addition to the polar localization of ColE1 in most cells, movement of the plasmid to the midcell position was observed in time-lapse studies. ColE1 could be displaced from its polar location by the p15A replicon, pBAD33, but not by plasmid RK2. The displacement of ColE1 by pBAD33 resulted in an almost random positioning of ColE1 foci in the cell and also in a loss of segregational stability, as evidenced by the large number of cells carrying pBAD33 with no visible ColE1 focus and as confirmed by ColE1 stability studies. The addition of the active partitioning systems of the F plasmid (sopABC) or RK2 (OB1 incC korB) resulted in movement of the ColE1 replicon from the cell pole to within the nucleoid region. This repositioning did not result in destabilization but did result in an increase in the number of plasmid foci, most likely due to partial declustering. These results are consistent with the importance of par regions to the localization of plasmids to specific regions of the cell and demonstrate both localization and dynamic movement for a naturally occurring plasmid that does not encode a replication initiation protein or a partitioning system that is required for plasmid stability.

scholarly journals Improved Cloning Vectors for Bifidobacteria, Based on the Bifidobacterium catenulatum pBC1 Replicon

2008 ◽  
Vol 74 (15) ◽  
pp. 4656-4665 ◽  
Author(s):  
Pablo Álvarez-Martín ◽  
Ana Belén Flórez ◽  
Abelardo Margolles ◽  
Gloria del Solar ◽  
Baltasar Mayo
Keyword(s):  
Bifidobacterium Longum ◽  
Antibiotic Resistance Genes ◽  
Tetracycline Resistance ◽  
Cloning Vectors ◽  
Relative Copy Number ◽  
E Coli ◽  
Segregational Stability ◽  
Genes Encoding ◽  
Peptide Gene ◽  
Multiple Cloning Sites

ABSTRACT This study reports the development of several cloning vectors for bifidobacteria based on the replicon of pBC1, a cryptic plasmid from Bifidobacterium catenulatum L48 thought to replicate via the theta mode. These vectors, in which antibiotic resistance genes encoding either erythromycin or tetracycline resistance acted as selection markers, were able to replicate in a series of eight Bifidobacterium species at frequencies ranging from 4.0 × 101 to 1.0 × 105 transformants μg−1 but not in Lactococcus lactis or Lactobacillus casei. They showed a relative copy number of around 30 molecules per chromosome equivalent and a good segregational stability, with more than 95% of the cells retaining the vectors after 80 to 100 generations in the absence of selection. Vectors contain multiple cloning sites of different lengths, and the lacZα peptide gene was introduced into one of the molecules, thus allowing the easy selection of colonies harboring recombinant plasmids in Escherichia coli. The functionality of the vectors for engineering Bifidobacterium strains was assessed by cloning and examining the expression of an α-l-arabinofuranosidase gene belonging to Bifidobacterium longum. E. coli and Bifidobacterium pseudocatenulatum recombinant clones were stable and showed an increase in α-arabinofuranosidase activity of over 100-fold compared to that of the untransformed hosts.

scholarly journals Effects of Three Different Nucleoid-Associated Proteins Encoded on IncP-7 Plasmid pCAR1 on Host Pseudomonas putida KT2440

2015 ◽  
Vol 81 (8) ◽  
pp. 2869-2880 ◽  
Author(s):  
Chiho Suzuki-Minakuchi ◽  
Ryusuke Hirotani ◽  
Masaki Shintani ◽  
Toshiharu Takeda ◽  
Yurika Takahashi ◽  
...  
Keyword(s):  
Pseudomonas Putida ◽  
Structural Instability ◽  
Metabolic Phenotype ◽  
Pseudomonas Putida Kt2440 ◽  
Host Chromosome ◽  
Content Type ◽  
Inorganic Ion ◽  
Segregational Stability ◽  
Genes Encoding ◽  
Associated Proteins

ABSTRACTNucleoid-associated proteins (NAPs), which fold bacterial DNA and influence gene transcription, are considered to be global transcriptional regulators of genes on both plasmids and the host chromosome. Incompatibility P-7 group plasmid pCAR1 carries genes encoding three NAPs: H-NS family protein Pmr, NdpA-like protein Pnd, and HU-like protein Phu. In this study, the effects of single or double disruption ofpmr,pnd, andphuwere assessed in hostPseudomonas putidaKT2440. Whenpmrandpndorpmrandphuwere simultaneously disrupted, both the segregational stability and the structural stability of pCAR1 were markedly decreased, suggesting that Pmr, Pnd, and Phu act as plasmid-stabilizing factors in addition to their established roles in replication and partition systems. The transfer frequency of pCAR1 was significantly decreased in these double mutants. The segregational and structural instability of pCAR1 in the double mutants was recovered by complementation ofpmr, whereas no recovery of transfer deficiency was observed. Comprehensive phenotype comparisons showed that the host metabolism of carbon compounds, which was reduced by pCAR1 carriage, was restored by disruption of the NAP gene(s). Transcriptome analyses of mutants indicated that transcription of genes for energy production, conversion, inorganic ion transport, and metabolism were commonly affected; however, how their products altered the phenotypes of mutants was not clear. The findings of this study indicated that Pmr, Pnd, and Phu act synergistically to affect pCAR1 replication, maintenance, and transfer, as well as to alter the host metabolic phenotype.

scholarly journals Resveratrol production in bioreactor: Assessment of cell physiological states and plasmid segregational stability

2015 ◽  
Vol 5 ◽  
pp. 7-13 ◽  
Author(s):  
Margarida S. Afonso ◽  
Susana Ferreira ◽  
Fernanda C. Domingues ◽  
Filomena Silva
Keyword(s):  
Segregational Stability

scholarly journals Replication and segregational stability of Bacillus plasmid pBAA1.

1989 ◽  
Vol 171 (2) ◽  
pp. 1166-1172 ◽  
Author(s):  
K M Devine ◽  
S T Hogan ◽  
D G Higgins ◽  
D J McConnell
Keyword(s):  
Segregational Stability
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