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 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.

Effect of sub-inhibitory concentrations of cefepime on biofilm formation by Pseudomonas aeruginosa

2021 ◽  
pp. 1-8
Author(s):  
Soheir A.A. Hagras ◽  
Alaa El-Dien M.S. Hosny ◽  
Omneya M. Helmy ◽  
Mounir M. Salem-Bekhit ◽  
Faiyaz Shakeel ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Clinical Isolates ◽  
Polymeric Chain ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Encoding Gene ◽  
Fimbrial Protein

This study investigated the effect of cefepime at sub-minimum inhibitory concentrations (sub-MICs) on in vitro biofilm formation (BF) by clinical isolates of Pseudomonas aeruginosa. The effect of cefepime at sub-MIC levels (½–1/256 MIC) on in vitro BF by six clinical isolates of P. aeruginosa was phenotypically assessed following 24 and 48 h of challenge using the tissue culture plate (TCP) assay. Quantitative real-time polymeric chain reaction (qRT-PCR) was employed to observe the change in expression of three biofilm-related genes, namely, a protease-encoding gene (lasA), fimbrial protein-encoding gene (cupA1), and alginate-encoding gene (algC), in a weak biofilm-producing strain of P. aeruginosa following 24 and 48 h of challenge with sub-MICs of cefepime. The BF morphology in response to cefepime was imaged using scanning electron microscopy (SEM). The TCP assay showed strain-, time-, and concentration-dependent changes in in vitro BF in P. aeruginosa following challenge with sub-MICs of cefepime, with a profound increase in strains with inherently no or weak biofilm-producing ability. RT-PCR revealed time-dependent upregulation in the expression of the investigated genes following challenge with ½ and ¼ MIC levels, as confirmed by SEM. Cefepime at sub-MICs could upregulate the expression of BF-related genes and enhance BF by P. aeruginosa clinical isolates.

A new cysteine-rich protein-encoding gene family in Giardia duodenalis

Gene ◽  
1996 ◽  
Vol 169 (1) ◽  
pp. 33-38 ◽  
Author(s):  
Nanhua Chen ◽  
Jacqueline A. Upcroft ◽  
Peter Upcroft
Keyword(s):  
Gene Family ◽  
Giardia Duodenalis ◽  
Cysteine Rich Protein ◽  
Protein Encoding ◽  
Encoding Gene

scholarly journals Dynamic Genome Editing Using In Vivo Synthesized Donor ssDNA in Escherichia coli

Cells ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 467
Author(s):  
Min Hao ◽  
Zhaoguan Wang ◽  
Hongyan Qiao ◽  
Peng Yin ◽  
Jianjun Qiao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genome Editing ◽  
Genome Engineering ◽  
Single Gene ◽  
Rolling Circle Replication ◽  
Rolling Circle ◽  
Cas9 Nuclease ◽  
Dynamic Genome ◽  
Circular Ssdna

As a key element of genome editing, donor DNA introduces the desired exogenous sequence while working with other crucial machinery such as CRISPR-Cas or recombinases. However, current methods for the delivery of donor DNA into cells are both inefficient and complicated. Here, we developed a new methodology that utilizes rolling circle replication and Cas9 mediated (RC-Cas-mediated) in vivo single strand DNA (ssDNA) synthesis. A single-gene rolling circle DNA replication system from Gram-negative bacteria was engineered to produce circular ssDNA from a Gram-positive parent plasmid at a designed sequence in Escherichia coli. Furthermore, it was demonstrated that the desired linear ssDNA fragment could be cut out using CRISPR-associated protein 9 (CRISPR-Cas9) nuclease and combined with lambda Red recombinase as donor for precise genome engineering. Various donor ssDNA fragments from hundreds to thousands of nucleotides in length were synthesized in E. coli cells, allowing successive genome editing in growing cells. We hope that this RC-Cas-mediated in vivo ssDNA on-site synthesis system will be widely adopted as a useful new tool for dynamic genome editing.

Steroid hormones acutely regulate expression of a nudix protein-encoding gene in the endometrial epithelium of sheep

2006 ◽  
Vol 73 (8) ◽  
pp. 967-976 ◽  
Author(s):  
Nancy H. Ing ◽  
Rebecca L. Wolfskill ◽  
Shauna Clark ◽  
Jennifer A. deGraauw ◽  
Clare A. Gill
Keyword(s):  
Steroid Hormones ◽  
Protein Encoding ◽  
Encoding Gene ◽  
Endometrial Epithelium

A putative cold shock protein-encoding gene isolated from Arthrobacter sp. A2-5 confers cold stress tolerance in yeast and plants

2014 ◽  
Vol 57 (6) ◽  
pp. 775-782 ◽  
Author(s):  
Seong-Kon Lee ◽  
Sung-Han Park ◽  
Jeong-Won Lee ◽  
Hae-Min Lim ◽  
Sun-Young Jung ◽  
...  
Keyword(s):  
Cold Stress ◽  
Stress Tolerance ◽  
Cold Shock ◽  
Cold Shock Protein ◽  
Protein Encoding ◽  
Encoding Gene
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