In conclusion, our study showed that the N-terminal domain of Pdc2p interacts

with the upstream region of THI genes and PDC5. In the mechanism for THI gene expression mediated by Pdc2p in response to thiamin starvation, not only the transactivation activity but also the recruitment to THI promoters seems to be enhanced via interaction with Thi3p (Fig. 4). It is highly likely that under thiamin-deprived conditions, the ternary Thi2p/Thi3p/Pdc2p complex is formed and transactivates THI genes in yeast cells. Conversely, the association of Pdc2p with PDC5 was unaffected by thiamin concentration in the medium. To date, a mechanism selleck chemicals llc underlying the regulation of PDC5 expression by TPP remains uncertain. As Pdc1p, a major isoform of yeast pyruvate decarboxylase, functions as a negative regulator for expression of PDC5 (Eberhardt et al., 1999), it will be interesting to investigate the relation between the TPP-binding of Pdc1p and the transcriptional control of PDC5. This work was supported in part by a research grant from the Vitamin B Research Committee of Japan. “
“Two bacterial strains involved

in syntrophic degradation of chloroacetamide herbicide butachlor were isolated from a rice paddy soil. Analysis of 16S rRNA gene sequences indicated that the two isolates were related to members of the genera Mycobacterium and Sphingobium, respectively. Thus, a pair consisted of Mycobacterium sp. J7A and Sphingobium sp. J7B could rapidly degrade butachlor (100 mg L−1) http://www.selleckchem.com/products/MK-1775.html at 28 °C within 24 h, while each isolate alone was not able to completely degrade butachlor. The isolate Mycobacterium sp. J7A was observed to grow slightly on butachlor, possibly utilizing the alkyl side chain of butachlor as its carbon and energy

source, but the isolate Sphingobium sp. J7B alone could not grow on 4��8C butachlor at all. Gas chromatography–mass spectrometry on catabolic intermediates revealed that the strain J7A produced and accumulated 2-chloro-N-(2,6-diethylphenyl) acetamide (CDEPA) during growth on butachlor. This intermediate was not further degraded by strain J7A, but strain J7B was observed to be able to completely degrade and grow on it through 2,6-diethylaniline (DEA). The results showed that butachlor was completely degraded by the two isolates by syntrophic metabolism, in which strain Mycobacterium sp. J7A degraded butachlor to CDEPA, which was subsequently degraded by strain Sphingobium sp. J7B through DEA. “
“Overlapping embedded genes, such as htgA/yaaW, are assumed to be rare in prokaryotes. In Escherichia coli O157:H7, gfp fusions of both promoter regions revealed activity and transcription start sites could be determined for both genes. Both htgA and yaaW were inactivated strand specifically by introducing a stop codon. Both mutants exhibited differential phenotypes in biofilm formation and metabolite levels in a nontargeted analysis, suggesting that both are functional despite YaaW but not HtgA could be expressed.

In conclusion, our study showed that the N-terminal domain of Pdc2p interacts

with the upstream region of THI genes and PDC5. In the mechanism for THI gene expression mediated by Pdc2p in response to thiamin starvation, not only the transactivation activity but also the recruitment to THI promoters seems to be enhanced via interaction with Thi3p (Fig. 4). It is highly likely that under thiamin-deprived conditions, the ternary Thi2p/Thi3p/Pdc2p complex is formed and transactivates THI genes in yeast cells. Conversely, the association of Pdc2p with PDC5 was unaffected by thiamin concentration in the medium. To date, a mechanism DAPT underlying the regulation of PDC5 expression by TPP remains uncertain. As Pdc1p, a major isoform of yeast pyruvate decarboxylase, functions as a negative regulator for expression of PDC5 (Eberhardt et al., 1999), it will be interesting to investigate the relation between the TPP-binding of Pdc1p and the transcriptional control of PDC5. This work was supported in part by a research grant from the Vitamin B Research Committee of Japan. “
“Two bacterial strains involved

in syntrophic degradation of chloroacetamide herbicide butachlor were isolated from a rice paddy soil. Analysis of 16S rRNA gene sequences indicated that the two isolates were related to members of the genera Mycobacterium and Sphingobium, respectively. Thus, a pair consisted of Mycobacterium sp. J7A and Sphingobium sp. J7B could rapidly degrade butachlor (100 mg L−1) PLX3397 mouse at 28 °C within 24 h, while each isolate alone was not able to completely degrade butachlor. The isolate Mycobacterium sp. J7A was observed to grow slightly on butachlor, possibly utilizing the alkyl side chain of butachlor as its carbon and energy

source, but the isolate Sphingobium sp. J7B alone could not grow on GNAT2 butachlor at all. Gas chromatography–mass spectrometry on catabolic intermediates revealed that the strain J7A produced and accumulated 2-chloro-N-(2,6-diethylphenyl) acetamide (CDEPA) during growth on butachlor. This intermediate was not further degraded by strain J7A, but strain J7B was observed to be able to completely degrade and grow on it through 2,6-diethylaniline (DEA). The results showed that butachlor was completely degraded by the two isolates by syntrophic metabolism, in which strain Mycobacterium sp. J7A degraded butachlor to CDEPA, which was subsequently degraded by strain Sphingobium sp. J7B through DEA. “
“Overlapping embedded genes, such as htgA/yaaW, are assumed to be rare in prokaryotes. In Escherichia coli O157:H7, gfp fusions of both promoter regions revealed activity and transcription start sites could be determined for both genes. Both htgA and yaaW were inactivated strand specifically by introducing a stop codon. Both mutants exhibited differential phenotypes in biofilm formation and metabolite levels in a nontargeted analysis, suggesting that both are functional despite YaaW but not HtgA could be expressed.

g. functional genomics, microarray analysis, immunochemical and infection model systems), appear to yield comprehensive and definitive information on protein function in fungi. The relative advantages of proteomic, as opposed to transcriptomic-only, analyses

are also described. In the future, combined high-throughput, quantitative proteomics, allied to transcriptomic sequencing, are set to reveal much about protein function in fungi. Fungal proteomics research, especially that related to filamentous fungi, has progressed dramatically over the past 5 years. This has been due to the availability of multiple fungal genome sequences, the advent of next-generation nucleic acid sequencing and the availability of powerful learn more proteomics technologies, especially tandem LC-MS (Martin et al., 2008; Braaksma et al., 2010; Costa et al., 2010). Combined, these technological advances have enabled high-throughput selleck chemical protein identification and functional assignment that was not even considered possible up to 10 years ago. The requirement to further understand the clinical consequences of opportunistic fungal infection, especially in immunocompromised patients, as well as the plant pathogenic nature of fungi, allied to the biotechnological potential of fungal enzymes for biofuel production, have also driven this intense activity (Taylor et

al., 2008; Dagenais & Keller, 2009; Schuster & Schmoll, 2010). Consequently, proteomics, by virtue of its capacity to yield definitive information on protein identity, localization, posttranslational modification and the accuracy of in silico gene model prediction in fungi, has become an integral component of all large-scale ‘omic’ and systems approaches to understanding the rich complexity of fungal biochemistry (Table 1). The lack of information that existed with respect to fungal proteomes has meant that

significant recent research has focused on Phloretin developing methodologies compatible with optimal protein extraction from fungi, and establishing basic data on the types and relative abundances of proteins present in fungi (Lakshman et al., 2008). Much effort has also been directed at cataloguing mycelial, organellar and secreted proteins (secretome) across a range of fungal species (Bouws et al., 2008; Kim et al., 2008). These approaches have used both individual protein identification following SDS-PAGE or 2D-PAGE fractionation or ‘shotgun’ proteomics, where total protein digests of fungal origin are analysed by tandem LC-MS to generate constituent protein data sets (Carberry et al., 2006; Braaksma et al., 2010). More recently, the dynamic nature of fungal proteomes has been investigated, whereby the effects of carbon sources, antifungal drugs and gene deletion have been explored at the proteomic level (Fernández-Acero et al., 2010; Cagas et al., 2011).

Ann Intern Med 2008; 148: 519–528. 71 Brook G, Main J, Nelson M et al. British HIV Association guidelines for the management

of coinfection with HIV-1 and hepatitis B or C virus 2010. HIV Med 2010; 11: 1–30. 72 Lubel JS, Angus PW. Hepatitis B reactivation in patients receiving cytotoxic chemotherapy: diagnosis and management. J Gastroenterol Hepatol 2010; 25: 864–871. 73 Davies JM, Lewis MP, Wimperis J et al. Review of guidelines for the prevention and treatment of infection in patients with an absent or dysfunctional spleen: prepared on behalf of the British Committee for Standards in Haematology by a working party of the Haemato-Oncology task force. Br J Haematol 2011; 155: 308–317. The writing group thanks the following for their comments and contributions to the guideline: Kirit Ardeshna Aravind Arumainathan Robin www.selleckchem.com/products/bmn-673.html Grant Sharon Jay Alistair Miller Josie Shew Lindsay Short Kate Templeton Laura Waters (on behalf of the BASHH HIV Special Interest Group) Prof Mark Bower has has received lecture fees, honoraria LGK 974 and advisory board attendance fees from Abbott, Bristol-Myers Squibb, Gallen, Gilead, Janssen & ViiV. Dr Adrian Palfreeman has no conflicts of interest to declare. Dr Maryam

Alfa-Wali has no conflicts of interest to declare. Prof Chris Bunker has no conflicts of interest to declare. Dr Fiona Burns has received speaker fees from Janssen and an educational travel grant from Gilead. Dr Duncan Churchill has, in the past year, received sponsorship from Janssen to attend a conference, and has sat on advisory boards for Gilead. Mr Simon Collins has no conflicts of interest to declare. Dr Kate Cwynarski has received advisory board honoraria/travel/registration reimbursement from Roche. Dr Simon Edwards has received

speaker, advisory and conference attendance fees from Merck Sharp and Dohme, Gilead, Abbott, ViiV and Janssen. Dr Paul Fields has no conflicts of interest to declare. Dr Kate Fife has no conflicts of interest to declare. Dr Eve Gallop-Evans has received ad board honoraria from Galen. Dr Shireen Kassam Phosphoprotein phosphatase has no conflicts of interest to declare. Dr Ranjababu Kulasegaram has received speaker and advisory fees from Merck Sharp and Dohme, Abbott, ViiV and Janssen. He has received research funding from Boehringer Ingelheim, Pfizer, ViiV and Gilead. He has received educational travel grants from Janssen, ViiV and Bristol-Myers Squibb. Prof Charles Lacey has received speaker fees from Sanofi Pasteur MSD Dr Robert Marcus has received lecture fees, honoraria and advisory board attendance fees from Roche and Napp Pharmaceuticals. Dr Silvia Montoto has no conflicts of interest to declare. Dr Mark Nelson has received lecture fees from Abbott, Boehringer Ingelheim, Bristol-Myers Squibb, Gilead, Merck Sharp & Dohme, Tibotec and ViiV and consultancy fees from Abbott, Boehringer Ingelheim, Bristol-Myers Squibb, Gilead, Idenix, Merck Sharp & Dohme, Pfizer, Tibotec and ViiV.

To eliminate the disturbing

effect of the fusion protein (Fig. 3b), the fusion transposase producer plasmid was eliminated from five yjjY mutants and the motility of these strains was tested again. Reduced motility was observed in all cases, indicating that in (or close to) the yjjY gene, a DNA segment is located that affects motility. Because the sequence of the yjjY insertion site showed high similarity to the consensus used by the wt IS30 transposase, we tested whether the wt IS30 uses this target sequence as a hot spot. Only seven yjjY mutants were Forskolin mw found to be generated by the wt IS30 out of the 222 mutants tested. These data demonstrate that the fusion transposase has a much more pronounced target preference for the yjjY hot spot (17.3%) compared with that of the wt transposase (3.2%). In this study, we have worked out and successfully applied a novel method based on IS30-mediated site-directed mutagenesis in order to produce nonflagellated S. Enteritidis mutants. The system was constructed based on the assumption that the FljA repressor component of the fusion transposase – as a DNA-binding protein – would bind to its target (the operator of fliC), and as a consequence, insertions could be concentrated with a relatively high frequency in the flagellin operon. The system constructed on the above basis worked well

and generated insertions. It turned out that the sequenced insertion sites showed pronounced similarity to the IS30 consensus sequence Rebamipide of insertions (Table 1;

Olasz et al., 1998). This Everolimus molecular weight indicated that the fusion transposase retained the target recognition ability of the wt IS30 transposase. Another feature of the insertions was that four target sites – called hot spots – were utilized several times. One of these hot spots was the target sequence in the fliD gene and these insertions resulted in nonmotile phenotypes. This fact could be considered as a proof of FljA-targeted transposition, because fliD is located in close proximity to the fliC operator sequence, which is the binding site of the native FljA repressor protein. These data suggested that the fusion of the FljA repressor protein modulated the target preference of the IS30 transposase and increased the frequency of integration into a new target site not preferred by the wt transposase. This result is in good agreement with earlier observations that the target preference of IS30 transposase can be modified by fusing the enzyme to unrelated DNA-binding proteins (Szabo et al., 2003 and unpublished data). Unexpectedly, another highly preferred hot spot was identified in the putative gene yjjY. Although this target site was recognized by both the wt and the fusion transposase, the frequency of the mutations generated by the IS30–FljA transposase was almost six times higher than that of the wild type (17.3% vs. 3.2%).

First, direct isolation and analysis of the end of the linear chromosome with its covalently attached terminal protein by biochemical means is definitive (Lin et al., 1993; Goshi et al., 2002). Secondly, an analysis of the gene topology by pulsed-field gel electrophoresis (PFGE) is highly suggestive (Rednenbach et al., 2000). Finally, identification of genes associated with chromosome linearity, such as tpg (gene encoding the terminal protein that is covalently linked to the end of the linear chromosome), tap (gene encoding a telomere-associated protein that seems to be essential to linear chromosome replication

and is usually closely linked with tpg on the chromosome) and ttr (gene encoding a protein U0126 mouse that is present very close to

ends of most linear chromosomes and seems to be involved in linear genome mobilization), implies linearity is present or was present at some point in the past (Goshi et al., 2002; Huang et al., 2007; Suzuki et al., 2008; Kirby & Chen, 2011). However, the absence of homologues of one or all of the tpg, tap and ttr trio does not confirm circularity because there is significant diversity in the terminal Selleck Anti-diabetic Compound Library replication mechanism of linear chromosomes and plasmids of Actinomycetales (Huang et al., 2007; Suzuki et al., 2008). The problems of defining linearity other than by definitive biochemical means, which is laborious, can be illustrated in a number of ways. Using PFGE, Saccharopolyspora erythraea NRRL 2338 was suggested BCKDHA to be linear based on analysis of the absence and presence of chromosome bands before and after proteinase

K treatment (Reeves et al., 1998). However, by chromosome sequencing, Oliynyk et al. (2007) indicated that the chromosome of this species is circular. Analysis at the gene level of the chromosome sequence does not identify any homologues of the tpg, tap and ttr trio or the presence of terminal repeats, which supports the latter conclusion. Notwithstanding the missed restriction sites pinpointed by the chromosome sequencing, the entry of the 8 Mb chromosome into the PFGE gel after proteinase K digestion, and the failure of the untreated chromosome to enter the gel under identical circumstances, supports directly the presence of bound terminal protein at the ends of a linear chromosome. Furthermore, Oliynyk et al. (2007) provide indirect evidence to support circularity, for example on the basis of the detection by gel electrophoresis of a fragment overlapping both proposed termini of the linear chromosome. The question remains somewhat open, but perhaps biased towards circularity. In the case of other Actinomycetales chromosome sequences, there is even less evidence to support circularity.

First, direct isolation and analysis of the end of the linear chromosome with its covalently attached terminal protein by biochemical means is definitive (Lin et al., 1993; Goshi et al., 2002). Secondly, an analysis of the gene topology by pulsed-field gel electrophoresis (PFGE) is highly suggestive (Rednenbach et al., 2000). Finally, identification of genes associated with chromosome linearity, such as tpg (gene encoding the terminal protein that is covalently linked to the end of the linear chromosome), tap (gene encoding a telomere-associated protein that seems to be essential to linear chromosome replication

and is usually closely linked with tpg on the chromosome) and ttr (gene encoding a protein check details that is present very close to

ends of most linear chromosomes and seems to be involved in linear genome mobilization), implies linearity is present or was present at some point in the past (Goshi et al., 2002; Huang et al., 2007; Suzuki et al., 2008; Kirby & Chen, 2011). However, the absence of homologues of one or all of the tpg, tap and ttr trio does not confirm circularity because there is significant diversity in the terminal GSK1120212 molecular weight replication mechanism of linear chromosomes and plasmids of Actinomycetales (Huang et al., 2007; Suzuki et al., 2008). The problems of defining linearity other than by definitive biochemical means, which is laborious, can be illustrated in a number of ways. Using PFGE, Saccharopolyspora erythraea NRRL 2338 was suggested Parvulin to be linear based on analysis of the absence and presence of chromosome bands before and after proteinase

K treatment (Reeves et al., 1998). However, by chromosome sequencing, Oliynyk et al. (2007) indicated that the chromosome of this species is circular. Analysis at the gene level of the chromosome sequence does not identify any homologues of the tpg, tap and ttr trio or the presence of terminal repeats, which supports the latter conclusion. Notwithstanding the missed restriction sites pinpointed by the chromosome sequencing, the entry of the 8 Mb chromosome into the PFGE gel after proteinase K digestion, and the failure of the untreated chromosome to enter the gel under identical circumstances, supports directly the presence of bound terminal protein at the ends of a linear chromosome. Furthermore, Oliynyk et al. (2007) provide indirect evidence to support circularity, for example on the basis of the detection by gel electrophoresis of a fragment overlapping both proposed termini of the linear chromosome. The question remains somewhat open, but perhaps biased towards circularity. In the case of other Actinomycetales chromosome sequences, there is even less evidence to support circularity.

, 2008). That is, because potato fields are commonly kept under slightly acidic conditions to avoid outbreaks of scab disease (Mizuno & Yoshida, 1993; Mishra & Srivastav, 1996; Lacey & Wilson, 2001), fungal antagonists would be expected to exert enhanced antagonistic activity under these conditions (Spadaro & Gullino, 2005). selleck Therefore, exploration of fungal antagonists is important not only for elucidating novel antagonistic functions of fungi but also for practical development of a method to biologically control potato scab disease. The bacterial strains used

in this study were obtained from JCM (Japan Collection of Microorganisms, Hirosawa, Wako, Japan). Streptomyces sp. were cultured on ISP medium 4 (Shirling & Gottlieb, 1966) selleck chemicals at 25 °C for 3 weeks to prepare spore suspensions for an antagonistic activity assay. Their CFUs were counted on GYM medium (glucose 4 g L−1, yeast extract 4 g L−1, malt extract 10 g L−1) solidified with 1.5% agar. Potato dextrose agar (PDA) (DSMZ medium129) and malt extract agar (malt extract 20 g L−1, glucose 20 g L−1, peptone 1 g L−1, agar 15 g L−1)

media, and one-tenth the strength of each of those media containing streptomycin (50 μg mL−1) and rose bengal (40 μg mL−1) were used to isolate fungi. The fungal strains were isolated from soils obtained from five potato fields in Abashiri, Hokkaido, Japan. Soil samples were serially diluted with sterile water, and see more 50 μL of the suspension was spread on the surface of the medium for isolation. After 2–5 days of incubation, >800 fungal colonies were randomly picked and were transferred to a fresh medium at least three times for purification. A fungal isolate of each group was used for an agar diffusion assay with S. turgidiscabiei. Fungal strains showing antagonistic activity in the assay were subsequently tested against S. scabiei and S. acidiscabiei. One-tenth strength of GYM medium

solidified with 1.0% agar was used for the agar diffusion assay. The medium pH was adjusted to 5.0 or 6.0. After autoclaving at 121 °C for 15 min, the medium was cooled to 40 °C in a water bath. Spores of each potato scab pathogen grown on plates of ISP medium 4 were scraped and suspended in sterile-distilled water, and were filtered with a 5.0 μm filter (Sartorius). To prepare the assay plates, an aliquot of spore suspension of each potato scab pathogen was added to a final concentration of 1.0 × 105 CFU mL−1, and 7 mL of GYM medium containing the spores was solidified in 60-mm Petri dishes. Fungal isolates were precultured on PDA plates, and tiny pieces of the agar containing fungal mycelia and conidia were inoculated at the center of the assay plates with a sterile needle. After 48 h of incubation at 25 °C, the diameter of the inhibition zone and that of the fungal colony were measured. The values of antagonistic activity by the fungi were calculated by subtraction of the fungal colony diameters from the inhibition zone diameters.

Morphine (10 mg/kg i.p.) reduced the ability of inhibitory synapses in midbrain slices to express LTPGABA both at 2 and 24 h after drug exposure but not after 5 days. Cocaine (15 mg/kg i.p.) impaired LTPGABA 24 h after exposure, but not at 2 h. Nicotine (0.5 mg/kg i.p.) impaired LTPGABA 2 h after exposure, but not after 24 h. Furthermore, LTPGABA was completely blocked 24 h following brief exposure to a stressful stimulus, a forced swim task. Our data suggest that drugs of abuse and stress trigger a common modification to inhibitory plasticity, synergizing with their collective effect at excitatory synapses.

Together, the net effect of addictive substances or stress is expected to increase excitability selleck inhibitor of VTA dopamine neurons, potentially contributing to the early stages of addiction. “
“It is unclear how a localized spinal cord injury may acutely affect locomotor networks of segments initially spared by the lesion. To investigate the process of secondary damage following spinal injury, we used the in

vitro model of the neonatal rat isolated spinal cord with transverse barriers at the low thoracic–upper lumbar region to allow focal application of kainate in hypoxic and aglycemic solution (with reactive oxygen species). The time-course and nature of changes in spinal locomotor networks downstream of the lesion site were investigated selleck chemicals llc over the first 24 h, with electrophysiological recordings monitoring fictive locomotion (alternating oscillations between flexor and extensor motor pools on either

side) and correlating any deficit with histological alterations. The toxic solution irreversibly suppressed synaptic transmission within Resminostat barriers without blocking spinal reflexes outside. This effect was focally associated with extensive white matter damage and ventral gray neuronal loss. Although cell losses were < 10% outside barriers, microglial activation with neuronal phagocytosis was detected. Downstream motor networks still generated locomotor activity 24 h later when stimulated with N-methyl-d-aspartate (NMDA) and serotonin, but not with repeated dorsal root stimuli. In the latter case, cumulative depolarization was recorded from ventral roots at a slower rate of rise, suggesting failure to recruit network premotoneurons. Our data indicate that, within the first 24 h of injury, locomotor networks below the lesion remained morphologically intact and functional when stimulated by NMDA and serotonin. Nevertheless, microglial activation and inability to produce locomotor patterns by dorsal afferent stimuli suggest important challenges to long-term network operation. "
“Humans and animals optimize their behavior by evaluating outcomes of individual actions and predicting how much reward the actions will yield. While the estimated values of actions guide choice behavior, the choices are also governed by other behavioral norms, such as rules and strategies.

However, plasmids are poorly understood in Xanthomonas spp. beyond the knowledge that they are often carriers of important virulence/avirulence genes (Vivian et al., 2001; Sundin, 2007), including avrBs1 (Stall et al., 1986; Swanson et al., 1988) and avrBs3/pthA (Bonas et al., 1989; Kim et al., 2006). Up to six avirulence genes were found clustered on a 90-kb plasmid in X. campestris pv. malvacearum strain Proteases inhibitor XcmH1005 (De Feyter & Gabriel, 1991). Plasmids in xanthomonads have been reported to carry determinants for resistance to copper or streptomycin (Stall et al., 1986; Minsavage et al., 1990), standard compounds used for bacterial plant disease control (McManus et al., 2002; Hopkins, 2004).

Indications of a 26.7-MDa plasmid were reported in the 1980s in strains of X. arboricola pv. pruni from the United States (Kado & Liu, 1981; Lazo & Gabriel, 1987; Randhawa & Civerolo, 1987), but further characterization of this plasmid stalled. We recently observed a similarly sized plasmid in the

European X. arboricola pv. pruni strain CFBP 5530. The objectives of this study were to sequence Dapagliflozin nmr and annotate this plasmid, conduct comparative genomic analysis against known Xanthomonas plasmids and complete chromosomal sequences, ascertain the prevalence among X. arboricola pv. pruni genotypes and determine whether it is unique to this pathovar, and thus may offer a means for identification at the pathovar level, discrimination that is not possible with currently available molecular diagnostic methods. Xanthomonas strains were routinely

grown on peptone yeast extract glycerol agar (NYGA) (Turner et al., 1984) and peptone yeast extract glycerol broth (NYGB) with incubation at 28 °C for 24–48 h. The presence of plasmid pXap41 was first confirmed in representative strains of X. arboricola pv. pruni with the plasmid profile determined after plasmid DNA extraction, as MRIP described in Zhou et al. (1990), and restriction with EcoRI (Fermentas SA, Mont-sur-Lausanne, Switzerland) according to the manufacturer’s instructions. Restriction products were then separated by electrophoresis on a 1% agarose gel containing ethidium bromide. For screening its presence in a larger number of strains, a pXap41-specific multiplex-PCR was established. For this purpose, primers targeting genes involved in pXap41 replication and mobilization were designed using the program fastpcr v5.4. A geographically and genetically representative collection of 35 X. arboricola pv. pruni isolates covering the full range of described genotypes (Zaccardelli et al., 1999; Boudon et al., 2005) and two strains each of six additional X. arboricola pathovars (Table 1) were screened for the presence of pXap41. The identity of all X. arboricola pv. pruni strains was confirmed using a duplex-PCR assay (Pothier et al., 2011) before screening for plasmid presence. Amplifications were carried out in a final volume of 20 μL using AccuStart PCR SuperMix (Quanta Biosciences, Gaithersburg, MD) and 0.2 μM of each primer.