An arsenate susceptibility test was performed with transformed and cultured Escherichia coli DH5α cells, which carried recombinant DNA of full-length arsenic (ars) operon, namely a putative membrane permease, ArsP; a transcriptional repressor, ArsR; an arsenate reductase, ArsC; and an arsenical-resistance membrane transporter, Acr3, from the Japanese urease-positive thermophilic Campylobacter lari (UPTC) CF89-12. The E. coli DH5α transformant showed reduced susceptibility to arsenate (~1536 μg/mL), compared to the control. Thus, these ars four-genes from the UPTC CF89-12 strain cells could confer a reduced susceptibility to arsenate in the transformed and E. coli DH5α cells. E. coli transformants with truncated ars operons, acr3 (acr3) and arsC-acr3 (∆arsC-acr3), of the ars operon, showed an MIC value of 384 μg/mL (~384 μg/mL), similar to the E. coli cells which carried the pGEM-T vector (control). Reverse transcription PCR confirmed in vivo transcription of recombinant full-length ars operon and deletion variants (∆acr3 and ∆arsC-acr3) in the transformed E. coli cells.

• MeSH
• arsen metabolismus   toxicita   MeSH
• Campylobacter lari účinky léků   genetika   metabolismus   MeSH
• Escherichia coli genetika   metabolismus   MeSH
• metabolické sítě a dráhy genetika   MeSH
• mikrobiální testy citlivosti MeSH
• operon * MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• stanovení celkové genové exprese MeSH
• transformace genetická MeSH
• Publikační typ
• časopisecké články MeSH

An arsenic (ars) four-gene operon, containing genes encoding a putative membrane permease (ArsP), a transcriptional repressor (ArsR), an arsenate reductase (ArsC) and an arsenical-resistance membrane transporter (Acr3) was first identified in urease-positive thermophilic Campylobacter (UPTC) isolate, CF89-12. UPTC CF89-12 and some other Campylobacter lari isolates contained their ars four-genes, similarly, differing from that in the reference C. lari RM2100 strain. Two putative promoters and a putative terminator were identified for the operon in UPTC CF89-12. In vivo transcription of the operon was confirmed in the UPTC cells. PCR experiments using two primer pairs designed in silico to amplify two arsR and arsC-acr3 segments, respectively, generated two amplicons, approximately 200 and 350 base pairs, with all 31 of 31 and 19 of 31 C. lari isolates (n = 17 for UPTC; n = 14 for UN C. lari), respectively. An inverted repeat forming a dyad structure, a potential binding site for a transcriptional repressor, was identified in the promoter region. Within the deduced 61 amino acids sequence of the putative arsR open reading frame from the UPTC CF89-12, a metal binding box and a DNA-binding helix-turn-helix motif were identified. The UPTC CF89-12 and some other UPTC isolates isolated from natural environment were resistant to arsenate.

• MeSH
• arsen * MeSH
• arsenát reduktasy genetika   MeSH
• bakteriální geny * MeSH
• bakteriální RNA genetika   MeSH
• Campylobacter lari genetika   izolace a purifikace   MeSH
• DNA bakterií genetika   MeSH
• DNA primery MeSH
• genetické lokusy MeSH
• konformace nukleové kyseliny MeSH
• molekulární sekvence - údaje MeSH
• motiv helix-turn-helix genetika   MeSH
• operon genetika   MeSH
• otevřené čtecí rámce MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• sekvence aminokyselin MeSH
• sekvenční analýza DNA MeSH
• sekvenční seřazení MeSH
• ureasa genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We aimed to clarify if the thermophilic Campylobacter lari organisms including urease-negative (UN) C. lari and urease-positive thermophilic Campylobacter (UPTC) can be differentiated at the species and/or subspecies levels by employing the full-length flaA gene and flaA short variable region (SVR) nucleotide sequence information or not. Thermophilic Campylobacter isolates (n = 45) including UN C. lari (n = 17), UPTC (n = 18), and Campylobacter jejuni (n = 10) were well discriminated at the isolate level by the unweighted pair group method using arithmetic means analysis and neighbor joining procedures constructed based on the full-length flaA gene and flaA SVR nucleotide sequence information. Thus, these procedures may possibly be useful for epidemimological studies for C. lari and C. jejuni.

• MeSH
• Campylobacter jejuni klasifikace   genetika   izolace a purifikace   MeSH
• Campylobacter lari klasifikace   genetika   izolace a purifikace   MeSH
• DNA bakterií chemie   genetika   MeSH
• flagelin genetika   MeSH
• genotyp MeSH
• lidé MeSH
• molekulární sekvence - údaje MeSH
• polymorfismus genetický MeSH
• sekvenční analýza DNA MeSH
• shluková analýza MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

When recombinant plasmid DNA from a genomic DNA library and inverse PCR products of Campylobacter sputorum biovar paraureolyticus LMG17591 strain were analyzed, an approximate 6.5-kb pair region, encoding a urease gene operon, was identified. Within the operon, seven closely spaced and putative open reading frames for ureG, ureH(D), ureA, ureB, ureC, ureE, and ureF were detected in order. A possible overlap was detected between ureG and ureH(D), ureH(D) and ureA, and ureE and ureF. In addition, two putative promoter structures, probable ribosome-binding sites and a putative ρ-independent transcriptional terminator structure were identified. The urease gene operon transcription in the cells was confirmed by the reverse transcription-PCR analysis. A neighbor-joining tree constructed based on the nucleotide sequence information of urease genes showed that C. sputorum biovar paraureolyticus formed a cluster with Arcobacter butzleri, urease-positive thermophilic Campylobacter and some Helicobacter spp., separating those from the other urease-producing bacteria, suggesting a commonly shared ancestry among these organisms.

• MeSH
• Campylobacter sputorum enzymologie   genetika   izolace a purifikace   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• genová knihovna MeSH
• molekulární sekvence - údaje MeSH
• operon MeSH
• otevřené čtecí rámce MeSH
• pořadí genů MeSH
• promotorové oblasti (genetika) MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie nukleových kyselin MeSH
• shluková analýza MeSH
• skot MeSH
• stanovení celkové genové exprese MeSH
• terminátorové oblasti (genetika) MeSH
• ureasa genetika   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Using two primer pairs constructed in silico for the amplification of the intervening sequences (IVSs) of the 23S rRNA gene sequences of the genus Taylorella, none of the three representative T. equigenitalis strains NCTC11184(T), Kentucky 188 and EQ59 was shown to contain any IVSs in the first quarter region. In the central region, all three strains possessed one approximately 70 bp IVS (TeIVS2) different from any IVSs found in T. asinigenitalis. The predicted secondary structure model of the IVSs contained stem and loop structures. The central region of the IVS-stem structure contains an identical double-stranded consensus 15-bp sequence. The purified RNA fraction from the three strains contained 16S and 4-5S RNA species but no 23S rRNA species. Thus, the primary 23S rRNA transcripts from the three strains would be cleaved into approximately 1.2- and 1.6-kb rRNA fragments and approximately 70-bp IVS. In addition, 16 other T. equigenitalis isolates were found to carry a similar 70-bp IVS in the central region and to produce fragmented 23S rRNA.

• MeSH
• bakteriální geny MeSH
• bakteriální RNA genetika   MeSH
• druhová specificita MeSH
• gramnegativní bakteriální infekce mikrobiologie   veterinární   MeSH
• introny genetika   MeSH
• koně MeSH
• konformace nukleové kyseliny MeSH
• konsenzuální sekvence MeSH
• molekulární sekvence - údaje MeSH
• nemoci koní mikrobiologie   MeSH
• posttranskripční úpravy RNA MeSH
• RNA ribozomální 23S genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční homologie nukleových kyselin MeSH
• sekvenční seřazení MeSH
• Taylorella equigenitalis genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

• MeSH
• Campylobacter coli genetika   izolace a purifikace   MeSH
• Campylobacter jejuni genetika   izolace a purifikace   MeSH
• flagelin genetika   MeSH
• molekulární sekvence - údaje MeSH
• otevřené čtecí rámce MeSH
• polymerázová řetězová reakce MeSH
• sekvence aminokyselin MeSH
• sekvenční analýza proteinů MeSH
• ureasa metabolismus   MeSH
• Geografické názvy
• Japonsko MeSH
• Severní Irsko MeSH