Construction of various ALA1-lexA or GRS1-lexA fusion constructs for the Western blot analyses was as previously

described [24]. Briefly, an initiator mutant of lexA was Luminespib chemical structure amplified by PCR as an SpeI-XhoI fragment and cloned in the pADH high-copy-number yeast shuttle vector. A wild-type (WT) or mutant ALA1 sequence containing base pairs -105 to -24 relative to ATG1 was amplified by PCR as a PstI-SpeI fragment and was cloned in-frame into the 5′ end of lexA, resulting in various ALA1-lexA fusion constructs. Construction of GRS1-lexA fusion constructs followed a similar strategy. The expression of these lexA fusion constructs was under the control of a constitutive ADH promoter [25]. The Western blot analysis was as previously described [24]. Complementation assays for the cytoplasmic and mitochondrial functions of ALA1 The yeast ALA1 knockout strain, TRY11 (MATa, his3Δ200, leu2Δ1, lys2-801, trp1Δ101, ura3-52, and ala1Δ::TRP1) 10058-F4 ic50 was maintained by a plasmid carrying the WT ALA1 gene

and a URA3 marker [26]. Complementation assays for the cytoplasmic function of plasmid-borne ALA1 and its derivatives were carried out by introducing a test plasmid (with a LEU2 marker) into TRY11 and determining the ability of transformants to grow in the presence of 5-fluoroorotic acid (5-FOA). Cultures were incubated at 30°C for 3~5 days or until colonies appeared. The transformants evicted the maintenance plasmid that carries the URA3 marker in the presence of 5-FOA. Thus, only an enzyme with cytoplasmic AlaRS activity encoded by the test plasmid could rescue the growth defect. Following 5-FOA selection, a single colony of transformants was selected and grown to the stationary phase in synthetic medium lacking leucine. Starting from a cell density of 1.0 A 600, cultures were 5-fold serially diluted, and 5-μl aliquots of each dilution were spotted onto the designated YPG plates. The plates were incubated at 30°C

for 3~5 days. Photos were taken of the complementation assays on day 3 following incubation. Because yeast cells cannot survive on glycerol without functional mitochondria, the transformants did not grow on YPG plates unless a functional mitochondrial AlaRS was generated by the test plasmid. Assays of the cytoplasmic and mitochondrial GlyRS activities DNA Damage inhibitor followed a similar protocol [21]. Reverse-transcription (RT)-PCR To determine the relative levels of specific ALA1-lexA mRNAs derived from the fusion constructs, a semiquantitative RT-PCR experiment was carried out following the protocols provided by the manufacturer (Invitrogen). Briefly, total RNA was first isolated from the transformants, and aliquots (~1 μg) of RNA were then reverse-transcribed into single-stranded complementary (c)DNA using an oligo-dT primer. After RNase H treatment, the single-stranded cDNA products were amplified by a PCR using a pair of specific primers.

Careful evaluation of adverse events is required as the drug is used more widely, particularly

monitoring for hepatotoxicity and cardiotoxicity. Pharmacological interactions must also be considered carefully. In light of the small number of available studies, bedaquiline should only be used in carefully monitored research settings. While this new drug may become a valuable player in the armamentarium used to tackle drug-resistant TB, its risks and benefits must first be better understood. Acknowledgments This project was supported by the National Health and Medical Research Council of Australia, APP1054107. Dr Menzies is the guarantor for this article, and takes responsibility for the integrity of the work as a whole. Conflict of interest Gregory J. Fox declares no conflict of interest. Dick Menzies declares no conflict of P505-15 order interest. P-gp inhibitor Open Access This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. References 1. World Health Organization. Global tuberculosis control 2012. Geneva: 2012. http://​www.​who.​int/​tb/​publications/​global_​report/​en/​. Accessed on

1 May 2013. 2. World Health Organization. Treatment of tuberculosis guidelines. Geneva: 2010. http://​www.​who.​int/​tb/​features_​archive/​new_​treatment_​guidelines_​may2010/​en/​index.​html. many Accessed on 1 May 2013. 3. Keshavjee S, Farmer PE. Tuberculosis, drug resistance, and the history of modern medicine. New Engl J Med. 2012;367:931–6.PubMedCrossRef 4. World Health Organization. Guidelines for the programmatic management of drug-resistant tuberculosis. Geneva 2011. http://​whqlibdoc.​who.​int/​publications/​2011/​9789241501583_​eng.​pdf. Accessed on 1 May 2013. 5. Ahuja SD, Ashkin D, Avendano M, et al. Multidrug resistant

pulmonary tuberculosis treatment regimens and patient outcomes: an individual patient data meta-analysis of 9,153 patients. PLoS Med. 2012;9:e1001300.PubMedCentralPubMedCrossRef 6. Orenstein EW, Basu S, Shah NS, et al. Treatment outcomes among patients with multidrug-resistant tuberculosis: systematic review and meta-analysis. Lancet Infect Dis. 2009;9:153–61.PubMedCrossRef 7. Johnston JC, Shahidi NC, Sadatsafavi M, Fitzgerald JM. Treatment outcomes of multidrug-resistant tuberculosis: a systematic review and meta-analysis. PLoS One. 2009;4:e6914.PubMedCentralPubMedCrossRef 8. Migliori GB, Sotgiu G, Gandhi NR, et al. The collaborative group for meta-analysis of individual patient data in MDR-TB. Drug resistance beyond XDR-TB: results from a large individual patient data meta-analysis. Eur Respir J. 2013;42:169–79.PubMedCrossRef 9. The Stop TB Partnership.

J Solid State Chem 2010, 183:901–908.CrossRef 17. Xu L, Song H, Chou L: Facile synthesis of nano-crystalline alpha-alumina

at low temperature via an absolute ethanol sol–gel strategy. Mater Chem Phys 2012, 132:1071–1076.CrossRef 18. Yu PC, Yang RG, Tsai YY, Sigmund W, Yen FS: Growth mechanism of single-crystal α-Al 2 O 3 nanofibers fabricated by electrospinning techniques. J Eur Ceram Soc 2011, 31:723–731.CrossRef 19. Kang W, Cheng B, Li Q, Zhuang X, Ren Y: A new method for preparing alumina nanofibers by electrospinning technology. Text Res J 2011,81(2):148–155.CrossRef 20. Chen Y, Liu S, Wang G: Kinetics and adsorption behavior SAR302503 of carboxymethyl starch on α-alumina in aqueous medium. J of Colloid and Interface Science

2006, 303:380–387.CrossRef 21. Ho YS, McKay G: Pseudo-second order model for sorption processes. Process Biochem 1999, 34:451–465.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions J-HK, S-JY, D-HK, H-JJ, T-YK, and K-HP participated in the material preparation Natural Product high throughput screening and data analysis. J-WL drafted the manuscript. All authors read and approved the final manuscript.”
“Background In the last two decades, tin dioxide (SnO2) has attracted a great interest because of its potential application for resistivity-type gas sensor devices. This is related to both high electric conductivity (approximately 102 Ω-1·cm-1), compatible with standard electronics, and to the fact that the surface is chemically very active, in the presence of oxidizing and reducing gases [1–3]. Among SnO2 solid state gas sensor devices, those employing thin film technology are the most promising

in terms of gas sensing response [4], stability, sensitivity, second and especially compatibility with the downscaling of the electronic devices [5, 6]. However, both thick and thin film performances are limited by the extension of active surface that potentially reduces their sensitivity. Nowadays, the research is focusing on nanostructured materials, like nanowires, nanorods, nanotubes, and nanoribbons [7, 8] because they have a large surface-to-volume ratio and show enhanced chemical stability [9, 10] and electrical performances [11]. Nanowires probably present the most interesting morphology for the fabrication of gas sensing devices, having about 30% atoms that are localized just at the surface, where the sensor transduction mechanism takes place [12, 13], and thus enhancing the sensitivity. This is why SnO2 nanowires seem to be an interesting active material for gas sensor nanometer-scaled devices. Another critical problem concerning the SnO2 thin films is the aging effect after their exposure to the surrounding atmosphere, which is related to undesired and uncontrolled adsorption of some contaminants at their surface, especially native oxide containing various C carbon species [14].

CNE1-LMP1 cells were treated with the small molecule inhibitor WHI-P131, a specific inhibitor of STAT3 phosphorylation at residue tyrosine 705 and serine

727. Both the promoter activity (Figure  4C) and the protein level (Figure  4D) of cyclin D1 decreased greatly upon WHI-P131 treatment. Treatment click here with PD98059, a chemical inhibitor that blocks the nuclear translocation of STAT3, also decreased cyclin D1 promoter activity (Figure  4C) and protein expression (Figure  4D). On the other hand, the data in Figure  4C and Figure  4D indicated that AG1478, an EGFR specific tyrosine kinase inhibitor, decreased the transcriptional activity of the cyclin D1 promoter and protein level. WHI-P131 was less efficient in the presence of PD98059 in cyclin D1 transcription (Figure  4C) but not cyclin D1 protein level

(Figure  4D). siSTAT3 or WHI-P131 induced a stronger inhibition of cyclin D1 promoter activity than siEGFR or AG1478. Taken together, these data Vorinostat suggest that both EGFR and STAT3 signaling pathways are involved in the transcriptional activity of Cyclin D1 promoter and protein levels. LMP1 regulated the nuclear EGFR and STAT3 binding to the cyclin D1 promoter region directly Next, we addressed whether the nuclear interaction of EGFR and STAT3 associates with the cyclin D1 promoter directly using electrophoresis mobility shift assay (EMSA) in CNE1 and CNE1-LMP1 cells. The probes, which contain EGFR or STAT3 binding sites according to the previous report [31], were labeled with biotin. As shown in Figure  5A, we found significant binding of nuclear protein to cyclin D1 (lane 2) while LMP1 promoted more nuclear protein binding (lane 3), indicating that LMP1 promoted STAT3 binding to the cyclin D1 promoter. The complex in CNE1-LMP1 cells was abolished by adding cold STAT3 binding sequence (Figure  5A, lane 4) but not by a mutation in the STAT3 binding

sequence (Figure  5A, lane 5) or a nonspecific binding sequence (Figure  5A, lane 6). After we mutated the plasmid containing functional mutated cyclin D1 promoters, we PRKACG could not detect the band in either CNE1 or CNE1-LMP1 cells (lanes 8 and 9 of Figure  5A). After the CNE1 cells were treated with IL-6 to induce STAT3 activation, we observed STAT3 binding in the cyclin D1 promoter (Figure  5B). After the CNE1-LMP1 cells were treated with the STAT3 inhibitors WHI-P131 and PD98059 (Figure  5B), we observed that STAT3 binding in the cyclin D1 promoter decreased. Taken together, LMP1 promoted STAT3 binding to the Cyclin D1 promoter. Figure 5 LMP1 increased the binding ability of transcription factors EGFR and STAT3 to cyclin D1 promoter in vitro . (A) STAT3 binding activities within the cyclin D1 promoter were examined by EMSA.

9 V) in both the anodic and cathodic scans, indicating that significant oxygen-containing species (e.g., hydroxyl) only form at higher potential, and therefore, the Au/Pd catalysts could remain active over a wider potential window without being poisoned by hydroxyl groups. This is further demonstrated by the chronoamperometry tests in Figure 3b. The Au25Pd and Au50Pd show the highest area-specific current density (normalized to the ECSA of Pd) initially and are able to maintain their superior stability even after 1 h at ca. 0.144 mA cm-2, which is significantly higher than that of the Pd black (0.0099 mA cm-2).

Durability of the Au/Pd NPs was evaluated under the AST protocol with potentials applied between 0.6V (5 s) and 0.95 V (5 s) up to 14,000 cycles. Figure 3c shows that the Au25Pd preserves https://www.selleckchem.com/products/CX-6258.html almost 90% of its initial ECSA in the first 7,000 cycles and 71% after 14,000 cycles. However, the ECSA loss for the Pd black is 35% in the first 7,000 cycles and 62% after 14,000 cycles. Not only the Au25Pd but also other Au/Pd catalysts demonstrate better electrochemical durability in the long-term AST. It is well known that dissolution of

Pd in acidic electrolytes starts from the formation of PdO or PdOH. As Figure 5a shows, Au25Pd can depress the adsorption of oxygen-containing species within the potential window during the cycling tests; therefore, 4SC-202 in vitro ensemble effect originated from the unique morphologies of the Au core in the Au25Pd may contribute to its superior durability. Conclusions We have demonstrated that by decreasing concentration of the Au solution, the oxyclozanide hollow Au cores in our unique Au/Pd core-shell NPs were formed with smaller crystalline grains and highly porous structures. Results indicated that these

Au/Pd catalysts show superior catalytic activities as ideal catalysts for formic acid oxidation. Furthermore, these Au/Pd catalysts show excellent electrochemical stability, CO oxidation ability and long-term durability. Particularly, the Au25Pd NPs synthesized in this study present the best catalytic properties due to their unique structure. The hollow and porous gold cores tuned by reduced Au concentrations in the core-shell structures may influence Pd distribution and morphologies on the Au core. These remarkable properties make the Au/Pd NPs the promising catalysts for DFAFCs. Acknowledgments This work was partially supported by the National Science Foundation (ECCS-0901849 and CMMI-1000831). References 1. Alden LR, Han DK, Matsumoto F, Abruña HD, DiSalvo FJ: Intermetallic PtPb nanoparticles prepared by sodium naphthalide reduction of metal-organic precursors: electrocatalytic oxidation of formic acid. Chem Mater 2006, 18:5591.CrossRef 2. Hoshi N, Kida K, Nakamura M, Nakada M, Osada K: Structural effects of electrochemical oxidation of formic acid on single crystal electrodes of palladium. J Phys Chem B 2006, 110:12480.CrossRef 3.

2010). The pyrenoid forming factor LCIB/C was found by the analysis of pmp1 and ad1 mutants

of C. reinhardtii, which are unable to grow at air-level CO2 but able to grow under very low CO2 conditions. Duanmu and Spalding (2011) tried to isolated suppressor mutants for pmp1 and ad1, which complement the “air-dying” phenotype of pmp1 and ad1, and successfully obtained four lines of mutants. From physiological analyses of photosynthetic parameters of these mutants, the complex modes of the CCM, which require or are independent of LCIB, were revealed. Such complex modes of the CCM in C. reinhardtii and in other P505-15 eukaryotic algae are tightly related to carbonic anhydrases (CAs), which selleck chemicals llc probably function as DIC-flow controllers at specific subcellular locations. Moroney et al. (2011) reviewed the possible functions of multiple subtypes of CAs in C. reinhardtii based upon their localizations and expression profiles. In the review, the occurrence of a cryptic component of extracellular CA, CAH8, which might be a critical component to form CO2 on the outside surface of the plasmalemma, was discussed. There were also two interesting hypotheses proposed in the review on the function

of stromal CA, CAH6 as a barrier to CO2 leaking from the chloroplast, and on the putative mitochondrial γ-CA moiety, which may be associated with the NADH dehydrogenase and Depsipeptide function as a CO2 converter analogous

to the cyanobacterial system. Mechanisms regulating the CCM in response to environmental CO2 are an intriguing aspect of this research field. Yamano et al. (2011) reported the function of the master regulator of CO2-responsive transcription of the CCM, in the green alga Volvox carteri, a multicellular alga closely related to C. reinhardtii indicated that Volvox possesses a CO2-inducible CCM. A putative master regulator gene for Volvox CCM, Volvox CCM1, was identified and sequence characteristics strongly suggested the function of this gene product is analogous to that in C. reinhardtii. CO2 may also affect physiological states other than CCM. Dillard et al. (2011) tested an effect of low CO2 acclimation on the cell-division cycle in C. reinhardtii and demonstrated that low CO2 treatment caused an apparent arrest of ongoing cell division and that the cells were transiently synchronized, thus revealing a potentially new aspect of CO2 response in eukaryotic algae. Baba et al. (2011) dissected the structure–function relationship of the promoter region of the H43/Fea1 protein gene, which is known to be stimulated at the transcriptional levels by both increments of pCO2 and iron limitation under cadmium enriched condition.

1996; White 1999; Draper et al. 2003). Therefore, we focus specifically on these geographic measures to develop our proposed local rarity ranking system. Classifying local rarity Based on our review of NatureServe’s and the IUCN’s systems, we establish a new local assessment level (L-rank) for categorizing

locally rare taxa within local jurisdictions and geographic regions. Under this proposed system, a taxon will be considered locally rare if it meets minimum P505-15 order area of occupancy levels using grids composed of 1 km × 1 km (1 km2) cells. Although grids composed of 2 km × 2 km cells are commonly used in factoring the G, N, and S ranks, data were available at a 1 km2 scale. Cells of this size create a more accurate picture and thereby alleviate some of the problems associated with models based on larger cell sizes (Thuiller et al. 2008). At the same time, 1 km2 cells are compatible with other commonly used metric grids (e.g., 1 ha or 100 km2 cells), thus simplifying conversion of data to other scales. Moreover, unlike global, national, or sub-national assessments, it is less prohibitive to collect local data at the 1 km2 scale within a reasonable amount of time and level of effort. Accordingly, the L-rank category is an incorporation and modification

of aspects of the NatureServe and IUCN systems find more and is specifically designed to be used in conjunction with NatureServe’s original geographic assessment scales. To identify and classify locally rare taxa through geographic analysis, we outline specific area of occupancy criteria to designate different levels of rarity at the local scale. While we lend our support to the IUCN’s explicit area of occupancy criteria for larger scales, the same numbers cannot be logically applied to local assessment levels due to the fact that many local jurisdictions are relatively small and have an overall area of <2,000 km2, the maximum range to be considered for conservation status many (IUCN 2001). If the IUCN’s area of occupancy criteria were applied to these

small jurisdictions, taxa distributed throughout the entire county would still meet the minimum criteria for conservation status at the local assessment level. Therefore, we created new area of occupancy criteria specifically for the local assessment level (Table 1). Numerical criteria were chosen qualitatively based upon analysis of criteria used by other systems, available information on average county sizes in the United States, and reviews of research showing the effects of range size on susceptibility to environmental and biological stressors. The “Critically Imperiled” range size criteria of 10 km2 used in our system is based directly on the IUCN criteria for “Critically Endangered” as it is a good measure of extreme rarity and vulnerability.

CISH and FISH analysis The CISH and FISH results were assessed using the categories proposed by Daniele et al. [18]. Four majors patterns were identified: balanced disomy (1.6-2.0 gene and chromosome 7 in all cells), balanced trisomy (2.2-3.0 gene and chromosome 7), balanced polysomy (3.1-4.4 gene and chromosome 7), low amplification (gene-to-chromosome 7 ratio 2.1-3.0), and high amplification (gene-to-chromosome 7 ratio > 3.0). We considered the presence of at least a group of 10 neoplastic cells showing gene gain as the positive cut off. The CISH and FISH signals were read

by 2 investigators (MM and ADB) independently from the results of the other assays. Statistical Analysis Agreements CRM1 inhibitor between the test results (IHC, CISH and FISH) were estimated using the Cohen’s k test and its relative 95% confidence interval (95% CI). Specificity, sensitivity, negative and positive predicted value (NPV and PPV, respectively), concordance and the 95% CI of the CISH assay were estimated considering the FISH result as the gold standard. Significance was assessed at 5% level. The statistical software package used for this analysis was SPSS for Windows (version 17.0; SPSS Inc., Chicago IL, USA). Results EGFR

gene copy number according to tumor histotype The CISH analysis was performed successfully on cell blocks of 20 NSCLC and 13 pulmonary mCRC. Of the 33 FNAC samples analyzed, 27 (82%) presented an increased EGFR GCN. In detail, as summarized in Table 1, 6 cases (18%) were disomic (1.6-2.0 balanced gene and chromosome Histone demethylase 7) (fig 1A, B), 10 (30%) presented Entospletinib low polysomy (trisomy: 2.2-3.0 balanced gene and chromosome 7) and 15 (45%) high polysomy (3.1-4.4 balanced gene and chromosome 7). The 2 amplified NCSLC (gene-to-chromosome 7 ratio ≥ 2), were 1 ADC and 1 LCC (fig 1C, D). No significant differences between NSCLC and pulmonary metastases from CRC, were observed in relation to the disomic or polysomic status. Table 1 Distribution of EGFR gene copy number evaluated by CISH according to tumor histotype Histotype N° of cases Disomy

Trisomy Polysomy Amplified ADC 7 1 2 3 1 LCC 8 2 1 4 1 SCC 5 1 3 1 0 mCRC 13 2 4 7 0 Total 33 6 10 15 2 ADC: adenocarcinoma; LCC: large cell carcinoma; SCC: squamous cell carcinoma; mCRC: metastatic colo-rectal cancer; Disomy: 1.6-2.0 balanced gene and chromosome 7; Trisomy: balanced 2.2-3.0 gene and chromosome 7; Polysomy: 3.1-4.4 balanced gene and chromosome 7; Amplified: gene-to-chromosome 7 ratio ≥ 2 Figure 1 EGFR CISH analysis on non small cell lung carcinoma. Two different patterns of gene and chromosome 7 copy number obtained by CISH on cell blocks prepared from two different Lung Carcinoma FNAC: (A) EGFR not amplified and (B) paired chromosome 7 disomy; (C) EGFR gene amplification with a clustered pattern and (D) trisomy of chromosome 7. Original magnification ×1000.

While the unfavorable endocrine effects of contest preparation APO866 solubility dmso have been documented in male bodybuilders [1, 2, 10], anecdotal reports from physique athletes also describe a state in which metabolic rate has slowed to an extent that exceeds the predicted magnitude, making weight loss increasingly difficult despite low caloric intakes and high training volumes. Although such reports could potentially be related to inaccurate dietary reporting [11, 12], these claims may be substantiated by a number of metabolic adaptations to weight loss, including adaptive thermogenesis [13–15], increased mitochondrial

efficiency [16–19], and hormonal alterations that favor decreased energy expenditure, decreased satiety, and increased hunger [1, 2, 10]. As a dieting phase progresses, such adaptations may threaten dietary adherence, make further weight loss increasingly difficult, and predispose the individual to rapid weight regain following the cessation of the diet. Although data documenting the attainment

and recovery from extreme changes in body composition is limited, the present article aims to investigate the condition of metabolic adaptation described by competitors and identify potential mechanisms to explain such a phenomenon. The endocrine response to an energy deficit A number of hormones play prominent roles in the regulation of body composition, energy intake, and energy expenditure. The hormones of the thyroid gland, particularly triiodothyronine DAPT mouse (T3), are known to play an important and direct role BCKDHA in regulating metabolic rate. Increases

in circulating thyroid hormones are associated with an increase in the metabolic rate, whereas lowered thyroid levels result in decreased thermogenesis and overall metabolic rate [20]. Leptin, synthesized primarily in adipocytes, functions as an indicator of both short and long-term energy availability; short-term energy restriction and lower body fat levels are associated with decreases in circulating leptin. Additionally, higher concentrations of leptin are associated with increased satiety and energy expenditure [21]. Insulin, which plays a crucial role in inhibiting muscle protein breakdown [22] and regulating macronutrient metabolism, is considered another “adiposity signal” [23]. Similar to leptin, high levels of insulin convey a message of energy availability and are associated with an anorexigenic effect. Conversely, the orexigenic hormone ghrelin functions to stimulate appetite and food intake, and has been shown to increase with fasting, and decrease after feeding [24]. Testosterone, known primarily for its role in increasing muscle protein synthesis and muscle mass [22], may also play a role in regulating adiposity [25]. Changes in fat mass have been inversely correlated with testosterone levels, and it has been suggested that testosterone may repress adipogenesis [25]. More research is needed to delineate the exact mechanism (s) by which testosterone affects adiposity.

The fact that pRet42a transfer is also decreased in a derivative lacking

the pSym of GR64 (GR64-5), points to a chromosomal location of the putative inhibitor locus. Similarly, S. fredii pSfr64a was unable to perform conjugative transfer or induce transfer of pSfr64b in R. etli genomic background (CFN2001-3). Only R. etli pRet42a was still able to induce pSfr64b transfer in the R. etli background (CFN2001-2). The pSym of GR64 differs from the typical R. etli pSym To further analyze the bean-nodulating S. fredii strain GR64, we performed a phylogenetic analysis with chromosomal genes (recA, rpoB), and with the plasmid-encoded genes nifH and repB. The results (Figure 4) show that, based on the phylogeny of the chromosomal genes, GR64 clusters within the fredii clade, while nifH

Saracatinib and repB genes group strain GR64 with other bean-nodulating Sinorhizobium strains isolated from the South of Spain (Granada and Sevilla) [22, 23] and from the North of Africa (Tunisia) [24] (Figure 4C). The data obtained indicate that GR64 has a S. fredii chromosome but carries a pSym that allows nodulation of Phaseolus. However, this plasmid differs from typical R. etli pSyms in its replication genes, allowing it to coexist with plasmid pSfr64a, which does share its replication genes with the R. etli pSym. Another feature ABT-263 in vitro that differentiates this pSym is the presence of a single copy of the nifH gene. Figure 4 Phylogeny of GBA3 S. fredii GR64. Maximum likelihood phylogenetic trees based on chromosomal: (A) recA, (B) rpoB, and plasmid: (C) nifH and (D) repB gene fragments. Arrows indicate the localization of S. fredii GR64, and R.etli CFN42. Discussion Genomic comparisons of S. meliloti, A. tumefaciens, and R. etli [25], and between Rhizobium

leguminosarum bv viciae and Rhizobium etli [26], have shown that chromosomes are well conserved both in gene content and gene order, whereas plasmids presented few common regions and lacked synteny, except for some pairs of plasmids whose features indicate that they were part of the ancestral genome, and may be considered as secondary chromosomes [26, 27]. In R. etli, the symbiotic and self-transmissible plasmids are the less conserved replicons [25] with fewer collinear blocks [26]. In this paper we show that a conjugative plasmid from a bean nodulating S. fredii strain is formed by large segments of replicons found in strains belonging to different species from diverse geographic origins. These replicons include two plasmids of R. etli, and a S. fredii chromosome. In GR64, bean-nodulation is provided by pSfr64b. Although the phylogenetic relationship of the GR64 nifH gene shows that it is closely related to the R. etli gene (Figure 4), pSfr64b differs from the typical R. etli pSym in other features (see above). We have previously reported that R.