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Our laboratory strain Yersinia enterocolitica strain WA-314 biogroup 1B serotype O:8 displayed a different adhesion behavior to host cells compared to other Yersinia strains. To investigate whether this is based on differences in the gene content of the large pYV virulence plasmid which contains the major Yersinia adhesin YadA, we set out to sequence pYV WA-314 . pYV WA-314 is very...
The pBR322 plasmid origin replication and oriC show similar responses to adenine methylation. Both are subject to sequestration by membrane fractions. In fact, like the host origin oriC, the RNA II promoter region of pBR322 is regulated by methylation of three GATC adenine methylation sites.The SeqA gene product acts in the negative control of oriC by sequestration. We suggest that the role of SeqA...
pHW126, pIGRK, pIGMS31 and pRAO1 are the only known members of a novel and as yet uncharacterised family of rolling circle plasmids. pHW126 contains only two open reading frames, of which one shows homology to pMV158-family mobilisation proteins. Here we provide evidence that the second open reading frame encodes a replication protein (Rep). Mutation or deletion of this gene resulted in replication...
The mercury(II)-resistance transposons Tn502 and Tn512 were sequenced and shown to be members of the Tn5053 family. They are currently the sole representatives from the clinical setting and were obtained from geographically disparate Pseudomonas aeruginosa strains. The family is comprised of six novel transposons that display genetic and structural variability that has arisen in different ways. The...
Using a variety of antibiotics, it was found that nine separate isolates of spontaneous antibiotic resistant mutants of Escherichia coli K12 pPSX-vioABCDE overproduce the anti-tumour antibiotic violacein. Subsequent analysis showed that seven of these mutations occurred on the plasmid pPSX-vioABCDE. The other two overproducing strains carried spontaneous chromosomal mutations to lincomycin and kanamycin...
A cryptic plasmid from Lactobacillus paracasei TXW isolated from koumiss, designated as pTXW, was sequenced and characterized. It is 3178bp in length with a G+C content of 42.9%. The plasmid pTXW was predicted to encode 4 putative ORFs. RepB shared high homology with initiator proteins of plasmids from the rolling circle replication (RCR) pMV158 family. copG was predicted to encode a transcriptional...
We describe here the construction of Gateway-compatible vectors, pBGP1-DEST and pPICZα-DEST, for rapid and convenient preparation of expression plasmids for production of secretory proteins in Pichia pastoris. Both vectors direct the synthesis of fusion proteins consisting of the N-terminal signal and pro-sequences of Saccharomyces cerevisiae α-factor, the recognition sites for Kex2 and Ste13 processing...
Using food and commensal lactic acid bacteria (LAB) as vehicles for DNA delivery into epithelial cells is a new strategy for vaccine delivery or gene therapy. However, present methods for DNA delivery with LAB have suffered low efficiency. Our goal was to develop a new system to deliver DNA into epithelial cells with high efficiency using food and commensal LAB. An Escherichia coli-LAB shuttle plasmid,...
Some strains of Serratia entomophila and S. proteamaculans cause amber disease of the New Zealand grass grub Costelytra zealandica (Coleoptera: Scarabaeidae), an important pasture pest in New Zealand. The disease determinants of S. entomophila, are encoded on a 153,404-bp plasmid, termed pADAP for amber disease associated plasmid. The S. proteamaculans strain 143 (Sp143) exhibits an unusual pathotype,...
Nocardia brasiliensis, is a bacteria that lives as saprophyte in soil and causes a disease called actinomycetoma in both human and animals. Nocardia brasiliensis is an intracellular, facultative bacterium that replicates and survives within host macrophages. The mechanisms involved in the evasion of the microbicidal actions of macrophages remain unclear. The filamentous growth of N. brasiliensis is...
Liver-specific gene knockdown cannot be achieved by short hairpin RNA (shRNA) generated by RNA polymerase III promoter. Here we constructed, modified and evaluated apolipoprotein A-I (ApoA-I) promoter-driven shRNA expression vectors against human telomerase reverse transcriptase (hTERT) in SMMC-7721 cells. The roles of the cis-acting hammerhead ribozyme and the specific pausing site MAZ, the liver-specific...
Corynebacterium renale harbors four small cryptic plasmids, pCR1, pCR2, pCR3 and pCR4, and can be a good system for understanding host–plasmid interactions. In the present study, effect of plasmid loss and their subsequent introduction on various properties of the host was evaluated. Loss of plasmids caused a reduction in bacterial size and also slowed down their growth rate, μ, and respiratory rate,...
pBTK445 is a newly described large (∼60Kb), low-copy number, conjugative plasmid indigenous to the sulfur-chemolithoautotroph Advenella kashmirensis. Based on its minimal replication region, a shuttle vector, pBTKS was constructed which can be used for diverse Alcaligenaceae members. The construct was found to be stably maintained both in the native host as well as in Escherichia coli in the absence...
Information transfer is fundamental to all life forms. In the third domain of life, the archaea, many of the genes functioning in these processes are similar to their eukaryotic counterparts, including DNA replication and repair, basal transcription, and translation genes, while many transcriptional regulators and the overall genome structure are more bacterial-like. Among halophilic (salt-loving)...
Streptomyces turgidiscabies Car8 is an actinobacterium that causes the economically important disease potato scab. Pathogenesis in this species is associated with a mobile pathogenicity island (PAISt) that site specifically inserts into the bacA gene in Streptomyces spp. Here we provide the 674,223bp sequence of PAISt, which consists of two non-overlapping modules of 105,364 and 568,859bp. These modules...
Strains ATCC 14028 and SL1344 of Salmonella enterica serovar Typhimurium are more virulent than LT2 in the BALB/c mouse model. Virulence plasmid swapping between strains ATCC 14208, LT2, and SL1344 does not alter their competitive indexes during mouse infection, indicating that the three plasmids are functionally equivalent, and that their contribution to virulence is independent from the host background...
This work sequenced and characterized a cryptic plasmid called pBSG3 from wild-type Bacillus amyloliquefaciens B3 – a powerful agent for suppression of plant pathogenic organisms. It is an 8439bp circular molecule, with G+C content of 40.3%. We provide evidence that pBSG3 replicates via the rolling-circle (RC) mechanism and, sequence comparisons place it in the pC194 family of rolling-circle-replicons...
The intergenic region linking conjugative transfer and replication copy control modules of IncF plasmids shows conservation of gene homology and organization. Genes distal to finO are coordinately expressed with the upstream transfer operon encoding the majority of conjugation genes in related plasmids. Here we investigate potential functions for these genes in copy number control and in processes...
Salmonella enterica serotype Choleraesuis (S. Choleraesuis) usually causes systemic infections in man and needs antimicrobial treatment. Multidrug resistance (MDR) in S. Choleraesuis is thus a great concern in the treatment of systemic non-typhoid salmonellosis. A large plasmid, pSC138, was identified in 2002 from a S. Choleraesuis strain SC-B67 that was resistant to all antimicrobial agents commonly...
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