Write my Paper for Cheap in High Quality -
Olmec Civilization - Ancient History Encyclopedia
Nov 12, 2017 Olmec time period, order custom essay online -
Olmec Civilization - CrystalinksAnalysis of the olmec period, mucR gene regulating biosynthesis of exopolysaccharides: implications for biofilm formation in voices in poetry, Sinorhizobium meliloti Rm1021. Departamento de Biología Molecular, Universidad Nacional de Río Cuarto, Córdoba, Argentina Search for more papers by this author. Departamento de Biología Molecular, Universidad Nacional de Río Cuarto, Córdoba, Argentina Search for more papers by this author. Editor: Yaacov Okon. Correspondence: Walter Giordano, Departamento de Biología Molecular, Universidad Nacional de Río Cuarto, 5800-Río Cuarto, Córdoba, Argentina. Tel.: +54 0358 467 6114; fax: +54 0358 467 6232; e-mail: firstname.lastname@example.org. Bacterial surface polysaccharides are crucial for establishment of successful rhizobia – legume symbiosis, and in most bacteria, are also critical for biofilm formation and surface colonization. In Sinorhizobium meliloti , the regulatory protein MucR controls exopolysaccharide production. To clarify the relationship between exopolysaccharide synthesis and olmec period, biofilm formation, we studied mucR expression under growth conditions that influence attachment to polyvinylchloride, developed a microtiter plate assay to quantify biofilm formation in S. meliloti strain Rm1021 and king, mutants defective in succinoglycan (EPS I) and/or galactoglucan (EPS II) production, and olmec time period, analyzed expression of C: The Potential Therapy for Hepatitis, EPS I and time period, EPS II genes by Essay, quantitative reverse transcriptase-PCR. Period? Consistent with previous studies of planktonic bacteria, we found that disruption of the mucR gene in Rm1021 biofilms increased EPS II, but reduced EPS I gene expression. mucR expression was not affected by environmental conditions that influence biofilm formation on polyvinylchloride, and year plan stalin, biofilm formation by time period, Rm1021 was independent of C, exopolysaccharide synthesis. Time Period? Other factors on the Rm1021 cell surface, and growth conditions, presumably regulate attachment and/or growth as a biofilm on polyvinylchloride. Rhizobia are soil bacteria with the capability to establish a symbiotic relationship with legume plants when soil nitrogen is king, limited. Rhizobial surface polysaccharides play important roles in symbiosis and olmec time, formation of active nodules. Mutants defective in the production of Potential Therapy for Hepatitis C, exopolysaccharides, lipopolysaccharides, and capsular polysaccharides usually show reduced induction of time, effective nodules, and king, are particularly affected in time period, the process of infection through infection threads (Hirsch, 1999). One of the helicopters in vietnam, best-studied exopolysaccharides produced by Sinorhizobium meliloti is succinoglycan (EPS I) (Reinhold et al. , 1994), which consists of repeated units of an octasaccharide containing one galactose and seven glucoses, and has characteristic succinyl, acetyl, and pyruvyl modifications. Time Period? A 25-kb region located in the second symbiotic megaplasmid (pSymB) in S. meliloti clusters the exo–exs genes necessary for the production of EPS I. The roles of Hepatitis Potential for Hepatitis, most of these genes have already been defined (Reuber & Walker, 1993). Sinorhizobium meliloti is also capable of producing a second exopolysaccharide known as galactoglucan (EPS II) (Her et al. , 1990; Zevenhuizen, 1997), which is synthesized under conditions of phosphate limitation (as often found in soils) (Zhan et al. , 1991; Mendrygal & González, 2000), in period, the presence of a mutation in the regulatory gene mucR (Zhan et al. , 1989; Keller et al. , 1995) or an intact copy of the transcriptional regulator expR (Glazebrook & Walker, 1989; Pellock et al. , 2002). EPS II is a polymer of helicopters in vietnam, disaccharide repeating units consisting of an acetylated glucose and a pyruvylated galactose (Her et al. , 1990). Time Period? A 32-kb cluster of helicopters in vietnam, genes (the exp genes) also located in pSymB is responsible for the production of olmec time, EPS II (Glazebrook & Walker, 1989). EPS I and EPS II are synthesized in two different fractions: high molecular weight (HMW) and low molecular weight (LMW). External addition of the LMW fractions of EPS I (trimers of the Promoting Business Ethics, octasaccharide), and time, oligomers (15–20 units of the disaccharide) of Essay Promoting, EPS II, can restore defective infection phenotypes in olmec time period, exopolysaccharide mutants, indicating that the example, establishment of symbiosis requires the time, presence of at least one of the LMW forms of either EPS I or EPS II (Battisti et al. Voices In Poetry? , 1992; González et al. , 1996). Bacterial surface components, such as exopolysaccharides, flagella, and lipopolysaccharides, are important not only in rhizobia – legume symbiosis but also in biofilm formation. Period? Biofilms are defined as microbial communities surrounded by a self-produced polymeric matrix and attached to a surface (Costerton et al. , 1995). The major components of biofilms are water (up to 97% of the total volume) and year plan, bacterial cells. The minor components are exopolysaccharides (Sutherland, 2004), proteins, DNA, and bacterial lysis products (Branda et al. Time? , 2005). C? Exopolysaccharides play important roles in olmec time, surface attachment and Essay about Business, development of time period, mature biofilms (Watnick & Kolter, 1999; Sutherland, 2001). The biofilm matrix provides bacteria with a physical barrier against antibiotics and defense compounds from the host (Gilbert et al. , 1997), and against various environmental stresses including UV radiation, pH changes, osmotic shock, and desiccation (Flemming, 1993). In S. meliloti , the regulatory protein MucR plays a key role in the control of C: The Therapy C, EPS I and EPS II production by binding to promoter regions in both exopolysaccharide biosynthesis gene clusters (Keller et al. , 1995; Bahlawane et al. Time Period? , 2008). About Business Ethics? A mutation in mucR results in the production of high levels of the time period, HMW fraction of EPS II, and the reduction of EPS I to trace levels (Zhan et al. , 1991; González et al. King? , 1996). MucR also causes feedback inhibition of its own transcription by binding to a short transcribed region located upstream of the coding region of olmec time period, mucR (Bertram-Drogatz et al. C: The Therapy Essay? , 1997). Rhizobia face a diversity of natural environments ranging from olmec time period a rhizosphere rich in nutrients and root exudates, to soils deficient in five year stalin, nitrogen, phosphorus, water, and/or other nutrients. The behaviors of biofilms on abiotic and biotic surfaces provide the basis for olmec time period, several survival strategies in helicopters used, bacteria, particularly nonspore formers such as rhizobia. Previous studies by our group suggest that biofilm formation in S. meliloti is time period, altered by Essay about Promoting, changes in environmental conditions and the nutritional status of the medium (Rinaudi et al. , 2006). Adhesion of olmec time, bacteria to different surfaces, and their self-aggregation, may be modulated by voices in poetry, regulation of exopolysaccharide synthesis. The present study is focused on the roles of transcriptional regulator mucR , and exopolysaccharide synthesis, in time period, biofilm formation by S. meliloti Rm1021. The strains used in this study are listed in Table 1. Mutations carried in Rm3131 (Keller et al. , 1995), Rm9020 (Glazebrook & Walker, 1991), and Rm10002 (Glazebrook & Walker, 1989) were transferred between S. meliloti strains by helicopters used, phage Φ M12 general transduction as described previously by Finan et al. Olmec Time? (1984). Rain Example? Antibiotics were added at the following concentrations: streptomycin, 500 μg mL −1 ; neomycin, 200 μg mL −1 ; tetracycline, 10 μg mL −1 ; oxytetracycline, 0.75 μg mL −1 ; and olmec, chloramphenicol, 20 μg mL −1 . Culture media and growth conditions. Sinorhizobium meliloti was grown in minimal Rhizobium defined medium (RDM) [5 g sucrose, 100 mL RDM A stock (6 g KNO 3 ; 1 g CaCl 2 ·2H 2 O; 2.5 g MgSO 4 ·7H 2 O; 1000 mL H 2 O), 100 mL RDM B stock (10 g K 2 HPO 4 ; 10 g KH 2 PO 4 ; 0.1 g FeCl 3 ·6H 2 O; 1000 mL H 2 O), 4 mL biotin stock (0.25 mg mL −1 ), 1 mL thiamine stock (10 mg mL −1 ), H 2 O q.s. to king shawshank 1000 mL] (Vincent, 1970), Luria–Bertani (LB) broth (Sambrook et al. Time Period? , 1989), or tryptone–yeast extract (TY) medium (Beringer, 1974) at 30 °C. Essay Promoting Business Ethics? RDM medium was supplemented when needed with 0.3 M sucrose, 0.15 M NaCl, 0.1–100 mM phosphate, or 7 mM CaCl 2 . Construction of the mucR::lacZ transcriptional fusion. A DNA region containing the mucR promoter was generated by PCR amplification of Rm1021 genomic DNA using the olmec time, following primers ( mucR sense, 5′-TTCGGGTCTGCTGC TCTTTG-3′; mucR antisense, 5′-ATGTGATCTGGTCGTC CTGC-3′) designed from the Essay about Business, S. Olmec Period? meliloti Rm2011 mucR sequence (Martín et al. , 2000). The PCR-amplified fragment was cloned upstream of a promoterless lacZ gene in helicopters used in vietnam, the wide-host-range vector pMP220 (Spaink et al. , 1987). The mucR::lacZ fusion plasmid was introduced by triparental mating into S. meliloti Rm1021. Bacterial liquid cultures comprising 10–15% of the flask volume were grown in olmec period, a rotary shaker (Model SI4-2 Shel Lab, 12-mm orbit, Sheldon Manufacturing Inc., OR) at 200 r.p.m. and at 30 °C for 72 h. Planktonic cells were removed from the flasks and biofilm rings growing on the glass in the interface between air and the culture medium were gently washed twice with a sterile physiological saline solution, collected in in poetry, an Eppendorf tube, centrifuged, and olmec time period, resuspended in cold Z-buffer [100 mM sodium phosphate (pH 7.0), 10 mM KCl, 1 mM MgSO 4 , 50 mM β-mercaptoethanol] for β-galactosidase activity assays, performed as described by shawshank, Miller (1972). β-Galactosidase activity in Miller units was calculated using the formula (1000 × OD 420 nm )/(OD 600 nm Δ T × V ), where Δ T is the olmec, reaction time (min) and V the initial volume of the culture used (mL). The biofilm formation assay, based on the method of O'Toole & Kolter (1998), relies on the ability of cells to adhere to Essay about Business Ethics the wells of period, 96-well microtiter dishes made of polyvinylchloride. To each well, 150 μL of a 1 : 100 dilution of an overnight culture (OD 600 nm 0.2) was added; the voices, plates were covered with plastic to olmec period prevent evaporation and incubated without agitation at 30 °C for 48 h. Planktonic cells were gently homogenized manually by repeated pipetting and Potential Therapy Essay, bacterial growth was quantified by measuring OD at 600 nm. Cultures were aspirated using an automatic hand pipette, and time period, wells were washed three times with 180 μL of sterile physiological saline solution and stained for 15 min with 150 μL of 0.1% crystal violet (CV). Each CV-stained well was then rinsed thoroughly and repeatedly with water, and scored for biofilm formation by addition of 150 μL 95% ethanol. The OD 560 nm of Hepatitis C: The Therapy, solubilized CV was determined using a MicroELISA Auto Reader (Series 700 Microplate Reader, Cambridge Technology). Biofilm rings from olmec 3-day-old S. meliloti cultures growing in RDM medium or RDM supplemented with either 0.3 M sucrose or 25 mM phosphate were gently washed twice with a sterile physiological saline solution, collected in an Eppendorf tube, centrifuged (10 000 g for 5 min at Essay about Ethics, 4 °C), and immediately used for RNA isolation. Total RNA was purified using the TRI ReagentLS kit (Cat # TS 120) following the manufacturer's protocol. Time Period? Samples were DNAse treated and stephen redemption, RNA was finally solubilized in RNAse-free water. Olmec Period? RNA concentrations were determined using a spectrophotometer at OD 260 nm . Real-time reverse transcriptase (RT)-PCR analysis. First-strand cDNA from two independent biological samples was generated using an AffinityScript QPCR cDNA Synthesis Kit (Stratagene, Cat #600559) with 1 μg total RNA per reaction. An aliquot (1 μL) of the about Business, cDNA reaction was used as a template for real-time PCR. The primer sequences used for real-time PCR were described by olmec time, Glenn et al. (2007). The probes and helicopters, oligonucleotide sequences used were: (DFAM, 6-carboxyfluorescein; HEX, hexachlorofluorescein; DBH1, Black Hole Quencher 1; DBH2, Black Hole Quencher 2). For real-time PCR analysis using SMc00128 or expE2 probes, Brilliant II QPCR master mix (Stratagene, Cat #600804) was used. For the olmec, SYBR Green protocol, the Brilliant SYBR ® Green QPCR Master Mix (Stratagene, Cat #600548) was used. The experiment was performed using the Mx3005P ™ Real-Time PCR System (Stratagene), programmed as follows: stage 1, 95 °C for voices, 120 s; stage 2, 95 °C for 15 s and 60 °C for 30 s (two-temperature cycle repeated 40 times). The expression of SMc00128 was used as an internal control for olmec period, normalization as described previously (Krol & Becker, 2004). A difference of one threshold cycle ( C T ) value equals a twofold change in gene expression. The fold change was calculated as , where C T is the level of gene expression in the specified strain. β-Galactosidase and biofilm formation assays were performed in five stalin, triplicate and repeated at least three times. Values were averaged, and the SDs were calculated. Data were subjected to one-way anova , followed by comparison of period, multiple treatment levels with the control, using post hoc Fisher's LSD test. All statistical analyses were performed using infostat software version 1.0. Effects of environmental conditions on mucR expression. MucR is a transcriptional repressor of the used in vietnam, exp genes involved in olmec time period, the biosynthesis of EPS II, and an activator of EPS I biosynthesis in S. Year Plan? meliloti (Keller et al. , 1995). Our previous studies showed that the ability of S. meliloti strain Rm1021 to attach and develop a biofilm on period polyvinylchloride is strongly affected by about Business, environmental conditions (Rinaudi et al. , 2006). To determine the role of MucR in this process, we studied the effect of various environmental conditions on mucR expression, using a mucR promoter fusion to the lacZ gene that encodes β-galactosidase. Growth of S. Olmec Time Period? meliloti in a nutritionally limiting environment is known to promote the acid rain is an of, transition from time a planktonic to a sessile mode of life. For example, Rm1021 forms a larger biofilm biomass when it is grown in five, a nutrient-poor RDM medium, as compared with enriched media such as LB or TY (Fujishige et al. , 2005). On the olmec time, other hand, similar levels of king, mucR expression were observed in olmec period, cells resuspended from 3-day-old biofilms grown in any of these three media (data not shown). We also studied mucR expression in biofilm cells grown in shawshank, RDM medium supplemented with 0.3 M sucrose, 0.15 M NaCl, 25 mM phosphate, or 7 mM calcium chloride, conditions that influence attachment to the polyvinylchloride surface (Rinaudi et al. , 2006). Time? Although biofilm formation was enhanced when any of these three components was added to used the medium, mucR expression was only slightly increased by addition of phosphate, and unaffected by addition of sucrose or calcium (Fig. 1). Addition of 0.15 M sodium chloride, which reduces biofilm formation, had no effect on reporter expression from the mucR promoter (Fig. Olmec Time Period? 1). These observations suggest that the ability of acid is an of, S. meliloti Rm1021 to sense nutritional and environmental conditions, with the olmec time, consequent transition from a planktonic to a sessile mode, and formation of Promoting Business Ethics, biofilms (Rinaudi et al. , 2006), is not mediated by changes in olmec time, mucR expression. Effect of Hepatitis C: The for Hepatitis Essay, RDM supplementation with various compounds on olmec time period mucR expression. For Hepatitis C? β-Galactosidase activity (expressed as Miller units) in planktonic and biofilm cells from Rm1021 mucR::lacZ grown for 3 days on RDM medium supplemented with 0.3 M sucrose, 0.15 M NaCl, 25 mM phosphate, and olmec time, 7 mM calcium chloride. Five Year Stalin? Assays were performed in triplicate, and the mean values and olmec time period, SDs are shown. Values having different letters are significantly different from helicopters used in vietnam each other according to Fisher's LSD test ( P <0.05). Because expression of the mucR promoter was slightly increased in the presence of 25 mM phosphate as compared with the regular RDM medium (12.5 mM phosphate) (Fig. 1), we evaluated mucR expression in biofilms from the Rm1021 mucR::lacZ strain under a range of olmec time, phosphate concentrations (0.1–100 mM). The increase in phosphate availability was correlated with increased β-galactosidase activity (Fig. 2). The presence of mucR is necessary for Essay about Business, EPS I production (Zhan et al. , 1991; Keller et al. Olmec Time? , 1995; Bertram-Drogatz et al. , 1998). EPS I production is dramatically enhanced at high phosphate concentrations (Mendrygal & González, 2000). In Poetry? Our results suggest that this enhancement is mediated by increased mucR expression. Effect of phosphate on mucR expression. β-Galactosidase activity (expressed as Miller units) in planktonic and biofilm cells from period Rm1021 mucR::lacZ grown for 3 days on voices RDM medium supplemented with phosphate at the indicated concentrations. Assays were performed in triplicate, and the mean values and SDs are shown. Olmec Time? Values having different letters are significantly different from each other according to Fisher's LSD test ( P <0.05). β-Galactosidase assays showed that mucR expression is maximal during the C: The C Essay, exponential phase of planktonic growth (OD 600 nm 0.8). Intermediate values of β-galactosidase activity were observed in olmec time period, the lag phase (OD 600 nm 0.2) and the stationary phase of voices in poetry, growth (OD 600 nm 1.2). The expression of mucR was lower in a 3-day-old biofilm than at any stage of growth (Fig. 3), consistent with the results described above. mucR expression in planktonic and biofilm cells. β-Galactosidase activity (expressed as Miller units) of olmec time period, planktonic cells from various stages in the growth curve, and of biofilm cells from Rm1021 mucR::lacZ grown for 3 days on RDM medium. Rain Is An? Assays were performed in olmec period, triplicate, and the mean values and SDs are shown. Values having different letters are significantly different from each other according to helicopters in vietnam Fisher's LSD test ( P <0.05). Effect of mucR, exoY , and expA mutations on attachment of strain Rm1021 to olmec period polyvinylchloride. To further elucidate the role of five year plan, MucR in biofilm development, attachment of a mucR mutant to olmec period polyvinylchloride wells was evaluated by CV staining. Biomass of 2-day-old biofilms of the helicopters in vietnam, mutant grown in RDM medium was not different from olmec time period that of wild-type Rm1021 (data not shown). Five Year Plan Stalin? Similar observations for these two strains were obtained in MGM medium with high (10 mM) and low (0.1 mM) phosphate (Rinaudi & González, 2009). The mucR mutant produces the HMW fraction of EPS II (González et al. , 1996), suggesting that the nonsymbiotically active fraction of EPS II of S. meliloti is not involved in attachment to polyvinylchloride under these conditions. To assess the olmec, contribution of EPS II and EPS I to biofilm formation of Rm1021 in RDM medium, we analyzed the polyvinylchloride attachment ability of exoY and expA mutants, which are defective in the biosynthesis of EPS I and EPS II, respectively. Biofilm biomass of both the mutants in RDM medium was similar to stephen shawshank that of Rm1021, indicating that these polysaccharides are not crucial for polyvinylchloride attachment under our conditions (Fig. Olmec? 4). An additional mutation in expA on the exoY mutant background did not result in a further decrease in biofilm formation (Fig. 4), which rules out the used in vietnam, possibility of a compensation effect on exopolysaccharide production. Bacterial growth and biofilm formation of static cultures of Sinorhizobium meliloti Rm1021 and mutants in time period, EPS I and/or EPS II. Biofilm was allowed to develop in RDM medium for 48 h, and then quantified. Assays were performed in triplicate, and the mean values and SDs are shown. Values having different letters are significantly different from each other according to Fisher's LSD test ( P <0.05). Expression of five plan, genes involved in the synthesis of exopolysaccharides in biofilms. Quantitative RT-PCR was used to evaluate the expression of genes involved in time period, the production of Promoting, EPS I and EPS II in biofilms of Rm1021 and olmec time period, its mucR mutant. Hepatitis C: The Therapy For Hepatitis? Expression of expE2 or exoY gene in biofilms grown in olmec period, RDM medium (0.015 M sucrose, 12.5 mM phosphate), and RDM supplemented with 0.3 M sucrose or 25 mM phosphate was analyzed as described in king shawshank redemption, M&M. Time? The expE2 gene encodes a glycosyltransferase involved in the synthesis of EPS II. The exoY gene encodes a galactosyltransferase responsible for shawshank, incorporation of the first galactose into the intermediate lipid in EPS I biosynthesis. Introduction of a mutation in the MucR regulator in Rm1021 led to increased transcription of the expE2 gene, relative to the wild type. Transcription of expE2 in biofilms formed by time period, the mucR mutant was enhanced by addition of 0.3 M sucrose to culture medium, but was reduced by a high phosphate concentration (Fig. 5). This is consistent with the findings that increased phosphate availability in planktonic bacteria blocks EPS II synthesis (Zhan et al. , 1991; Mendrygal & González, 2000), and thereby reduces expression of the genes responsible for EPS II production. Acid Rain? Because expE2 is actively transcribed in biofilms of Rm1021 mucR (a strain that produces HMW EPS II) grown in RDM medium, and biofilm formation in Rm1021 mucR is similar to olmec time period that in the wild type (present study, and Rinaudi & González, 2009), the above finding confirms that the HMW fraction of EPS II produced by of, the mucR mutant is olmec time period, not involved in biofilm formation. Expression of the expE2 and exoY genes in voices, Sinorhizobium meliloti biofilms. Negative change values indicate downregulation in the mutant strain (Rm1021 mucR ) compared with expression in wild-type Rm1021. Results are means from two independent experiments. Values having different letters are significantly different from olmec period each other according to in poetry Fisher's LSD test ( P <0.05). exoY expression in biofilms of the mucR mutant was less than that in Rm1021 (Fig. 5). This result is consistent with previous observations that MucR promotes EPS I synthesis in planktonic bacteria (Bertram-Drogatz et al. , 1998). On the other hand, exoY expression was not activated by period, 25 mM phosphate (Fig. 5), suggesting that higher concentrations of phosphate are needed for induction of voices, EPS I production. Mendrygal & González (2000) reported that S. meliloti achieves the olmec time, maximal production of EPS I at phosphate concentrations higher than those used in the present study. In conclusion, our findings suggest that in vitro polyvinylchloride attachment by Rm1021 does not depend on exopolysaccharide synthesis under our experimental conditions. In contrast, Fujishige et al. (2006) found that succinoglycan (EPS I) is Essay about Ethics, involved in biofilm development. This apparent discrepancy may be explained by the fact that sucrose concentration in RDM medium for S. meliloti growth was 2% in the Fujishige study, but only 0.5% in the present study. High levels of sucrose in culture medium have been reported to time period cause increased exopolysaccharide synthesis in used, other microorganisms (van Geel-Schutten et al. , 1998; Lee et al. Olmec Period? , 2003; Gross & Rudolph, 2008), probably as a result of acid rain example, facilitated carbon uptake. Under our assay conditions, mucR gene expression and time period, regulation of five year stalin, exopolysaccharide biosynthesis do not appear to be crucial for olmec time period, biofilm formation in S. meliloti strain Rm1021. This finding is important for Hepatitis C: The Therapy C Essay, understanding the contribution of rhizobial exopolysaccharides to legume colonization, a key step in the nodulation pathway. This paper was written in partial fulfillment of the PhD thesis of L.V.R. to olmec time period the Departamento de Biología Molecular, Universidad Nacional de Río Cuarto (UNRC). L.V.R. and F.S. were supported by a fellowship from the Consejo Nacional de Investigaciones Científicas y Técnicas of the República Argentina (CONICET). This work was supported by grants from the Secretaría de Ciencia y Técnica de la UNRC, Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT), and CONICET. W.G. and A.Z. are Career Members of helicopters, CONICET. We are grateful to Drs A. Pühler and time period, G. Hepatitis Therapy? Walker for strains and Dr S. Anderson for editing the manuscript. © 2009 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. Olmec? All rights reserved. rhizobia; mucR ; exopolysaccharides; biofilms. Issue online: 27 November 2009 Version of Essay about Promoting Ethics, record online: 22 October 2009 Received 7 October 2009; accepted 9 October 2009.Final version published online November 2009. Bahlawane C , McIntosh M , Krol E & Becker A ( 2008 ) Sinorhizobium meliloti regulator MucR couples exopolysaccharide synthesis and motility . Mol Plant Microbe In 21 : 1498 – 1509 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 7 Battisti L , Lara JC & Leigh JA ( 1992 ) Specific oligosaccharide form of the Rhizobium meliloti exopolysaccharide promotes nodule invasion in alfalfa . P Natl Acad Sci USA 89 : 5625 – 5629 . CrossRef | PubMed | CAS | Web of time period, Science® Times Cited: 117 | ADS Beringer JE ( 1974 ) R factor transfer in Rhizobium leguminosarum . J Gen Microbiol 84 : 188 – 198 . CrossRef | PubMed | CAS | Web of five plan stalin, Science® Times Cited: 1227 Bertram-Drogatz PA , Rüberg S & Becker A ( 1997 ) The regulatory protein MucR binds to a short DNA region located upstream of the mucR coding region in olmec time period, Rhizobium meliloti . Mol Gen Genet 254 : 529 – 538 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 21 Bertram-Drogatz PA , Quester I , Becker A & Pühler A ( 1998 ) The Sinorhizobium meliloti MucR protein, which is essential for the production of Essay about Promoting Ethics, high-molecular-weight succinoglycan exopolysaccharide, binds to olmec time short DNA regions upstream of exoH and exoY . Mol Gen Genet 257 : 433 – 441 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 24 Branda SS , Vik S , Friedman L & Kolter R ( 2005 ) Biofilms : the matrix revisited . Trends Microbiol 13 : 20 – 26 . PubMed | CAS | Web of Science® Times Cited: 205 Costerton JW , Lewandowski Z , Caldwell DE , Korber DR & Lappin-Scott HM ( 1995 ) Microbial biofilms . Annu Rev Microbiol 49 : 711 – 745 . Stephen Shawshank Redemption? CrossRef | PubMed | CAS | Web of Science® Times Cited: 1373 Finan TM , Hartweig E , LeMieux K , Bergman K , Walker GC & Signer ER ( 1984 ) General transduction in Rhizobium meliloti . Period? J Bacteriol 159 : 120 – 124 . PubMed | CAS | Web of Science® Times Cited: 169 Finan TM , Kunkel B , De Vos GF & Signer ER ( 1986 ) Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and helicopters, thiamine synthesis genes . J Bacteriol 167 : 66 – 72 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 337 Flemming H-C ( 1993 ) Biofilms and environmental protection . Time Period? Water Sci Technol 27 : 1 – 10 . Voices? CAS | Web of Science® Times Cited: 36 Fujishige NA , Rinaudi L , Giordano W & Hirsch AM ( 2005 ) Superficial liaisons : colonization of roots and abiotic surfaces by rhizobia. Biology of Plant–Microbe Interactions. Olmec Time? Proceedings of the 12th International Congress on Molecular Plant–Microbe Interactions , Vol. Voices In Poetry? 5 ( SánchezF , QuintoC , López-LaraIM & GeigerO , eds), pp. 292 – 299 . Olmec? St. Paul, MN. Fujishige NA , Kapadia NN , De Hoff PL & Hirsch AM ( 2006 ) Investigations of Rhizobium biofilm formation . Rain Is An Of? FEMS Microbiol Ecol 56 : 195 – 206 . Wiley Online Library | PubMed | CAS | Web of Science® Times Cited: 31 Gilbert P , Das J & Foley I ( 1997 ) Biofilms susceptibility to antimicrobials . Adv Dent Res 11 : 160 – 167 . CrossRef | PubMed | CAS Glazebrook J & Walker GC ( 1989 ) A novel exopolysaccharide can function in place of the calcofluor-binding exopolysaccharide in nodulation of alfalfa by Rhizobium meliloti . Cell 56 : 661 – 672 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 179 Glazebrook J & Walker GC ( 1991 ) Genetic techniques in time period, Rhizobium meliloti . Method Enzymol 204 : 398 – 418 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 79 Glenn SA , Gurich N , Feeney MA & González JE ( 2007 ) The ExpR/Sin quorum-sensing system controls succinoglycan production in Sinorhizobium meliloti . J Bacteriol 189 : 7077 – 7088 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 13 González JE , Reuhs BL & Walker GC ( 1996 ) Low molecular weight EPS II of Rhizobium meliloti allows nodule invasion in Medicago sativa . Hepatitis C: The C Essay? P Natl Acad Sci USA 93 : 8636 – 8641 . CrossRef | PubMed | Web of Science® Times Cited: 95 | ADS Gross M & Rudolph K ( 2008 ) Studies on the extracellular polysaccharides (EPS) produced in olmec time period, vitro by Pseudomonas phaseolicola . III. Year Stalin? Kinetics of olmec, levan and alginate formation in batch culture and demonstration of levansucrase activity in crude EPS . J Phytopathol 119 : 289 – 297 . Wiley Online Library | Web of Science® Times Cited: 13 Her GR , Glazebrook J , Walker GC & Reinhold VN ( 1990 ) Structural studies of acid is an, a novel exopolysaccharide produced by time period, a mutant of Rhizobium meliloti strain Rm1021 . Carbohyd Res 198 : 305 – 312 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 49 Hirsch AM ( 1999 ) Role of lectins (and rhizobial exopolysaccharides) in C: The Potential Therapy for Hepatitis Essay, legume nodulation . Curr Opin Plant Biol 2 : 320 – 326 . Olmec Time Period? CrossRef | PubMed | CAS | Web of Science® Times Cited: 81 Keller M , Roxlau A , Weng WM , Schmidt M , Quandt J , Niehaus K , Jording D , Arnold W & Pühler A ( 1995 ) Molecular analysis of the Rhizobium meliloti mucR gene regulating the biosynthesis of the exopolysaccharides succinoglycan and galactoglucan . Mol Plant Microbe In 8 : 267 – 277 . CrossRef | CAS | Web of Science® Times Cited: 60 Krol E & Becker A ( 2004 ) Global transcriptional analysis of the year plan, phosphate starvation response in Sinorhizobium meliloti strains 1021 and 2011 . Mol Genet Genomics 272 : 1 – 17 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 62 Lee S , Dong-Hyuk S , Hey-Lin P , Choi Y & Jung S ( 2003 ) Solubility enhancement of a hydrophobic flavonoid, luteolin by the complexation with cyclosophoraoses isolated from Rhizobium meliloti . Time Period? Antonie van Leeuwenhoek 84 : 201 – 207 . Hepatitis Potential Therapy? CrossRef | PubMed | CAS | Web of Science® Times Cited: 15 Leigh JA , Signer ER & Walker GC ( 1985 ) Exopolysaccharide-deficient mutants of Rhizobium meliloti that form ineffective nodules . P Natl Acad Sci USA 82 : 6231 – 6235 . CrossRef | PubMed | CAS | Web of olmec time, Science® Times Cited: 394 | ADS Martín M , Lloret J , Sánchez-Contreras M , Bonilla I & Rivilla R ( 2000 ) MucR is about Business, necessary for galactoglucan production in Sinorhizobium meliloti EFB1 . Mol Plant Microbe In 13 : 129 – 135 . Time? CrossRef | PubMed | Web of Science® Times Cited: 34 Mendrygal KE & González JE ( 2000 ) Environmental regulation of exopolysaccharide production in helicopters used in vietnam, Sinorhizobium meliloti . Period? J Bacteriol 182 : 599 – 606 . Five Plan Stalin? CrossRef | PubMed | CAS | Web of Science® Times Cited: 46 Miller JH ( 1972 ) Experiments in Molecular Genetics . Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. O'Toole GA & Kolter R ( 1998 ) Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways : a genetic analysis . Mol Microbiol 28 : 449 – 461 . PubMed | CAS | Web of Science® Times Cited: 589 Pellock BJ , Teplitski M , Boinay RP , Bauer WD & Walker GC ( 2002 ) A LuxR homolog controls production of symbiotically active extracellular polysaccharide II by time, Sinorhizobium meliloti . J Bacteriol 184 : 5067 – 5076 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 59 Reinhold BB , Chan SY , Reuber TL , Marra A , Walker GC & Reinhold VN ( 1994 ) Detailed structural characterization of in poetry, succinoglycan, the period, major exopolysaccharide of redemption, Rhizobium meliloti Rm1021 . J Bacteriol 176 : 1997 – 2002 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 90 Reuber TL & Walker GC ( 1993 ) Biosynthesis of succinoglycan, a symbiotically important exopolysaccharide of Rhizobium meliloti . Cell 74 : 269 – 280 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 160 Rinaudi L , Fujishige NA , Hirsch AM , Banchio E , Zorreguieta A & Giordano W ( 2006 ) Effects of olmec, nutritional and environmental conditions on Sinorhizobium meliloti biofilm formation . Res Microbiol 157 : 867 – 875 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 12 Rinaudi LV & González JE ( 2009 ) The low-molecular-weight fraction of the exopolysaccharide II from Sinorhizobium meliloti is in poetry, a crucial determinant of biofilm formation . Olmec Time? J Bacteriol 191 : 7216 – 7224 . CrossRef | PubMed | CAS | Web of helicopters in vietnam, Science® Times Cited: 2 Sambrook J , Fritsch EF & Maniatis F ( 1989 ) Molecular Cloning: A Laboratory Manual , 2nd edn . Cold Spring Harbor Laboratory, Cold Spring Harbour, New York. Spaink HP , Okker RJH , Wijffelman CA , Pees E & Lugtenberg JJ ( 1987 ) Promoters in the nodulation region of the Rhizobium leguminosarum Sym plasmid pRL1J1 . Plant Mol Biol 9 : 27 – 39 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 429 Sutherland IW ( 2001 ) Exopolysaccharides in biofilms, flocs and related structures . Water Sci Technol 43 : 77 – 86 . Time Period? PubMed | CAS | Web of Science® Times Cited: 45 Sutherland IW ( 2004 ) Microbial exopolysaccharides . Voices? Polysaccharides: Structural Diversity and Functional Versatility , 2nd edn ( DumitriuS , ed), pp. 431 – 458 . Marcel Dekker, New York. Van Geel-Schutten GH , Flesch F , Ten Brink B , Smith MR & Dijkhuizen L ( 1998 ) Screening and characterization of Lactobacillus strains producing large amounts of exopolysaccharides . Appl Microbiol Biot 50 : 697 – 703 . CrossRef | CAS | Web of Science® Times Cited: 39 Vincent JM ( 1970 ) A Manual for the Practical Study of the Root of olmec time period, Nodule-Bacteria . Blackwell Scientific Publications, Oxford, UK. Watnick PI & Kolter R ( 1999 ) Steps in the development of a Vibrio cholerae El Tor biofilm . Year Stalin? Mol Microbiol 34 : 586 – 595 . Period? Wiley Online Library | PubMed | CAS | Web of Science® Times Cited: 234 Zevenhuizen LPTM ( 1997 ) Succinoglycan and galactoglucan . Essay About Business Ethics? Carbohyd Polym 33 : 139 – 144 . CrossRef | CAS | Web of Science® Times Cited: 10 Zhan HJ , Levery SB , Lee CC & Leigh JA ( 1989 ) A second exopolysaccharide of time, Rhizobium meliloti strain SU47 that can function in root nodule invasion . P Natl Acad Sci USA 86 : 3055 – 3059 . CrossRef | PubMed | CAS | Web of Science® Times Cited: 90 | ADS Zhan HJ , Lee CC & Leigh JA ( 1991 ) Induction of the second exopolysaccharide (EPSb) in Rhizobium meliloti SU47 by low phosphate concentrations . J Bacteriol 173 : 7391 – 7394 . Five Year Plan? CrossRef | PubMed | CAS | Web of Science® Times Cited: 48. Articles related to the one you are viewing. 1 J Nogales , D Pérez-Mendoza , M Gallegos , M Soto , Beneficial Plant-microbial Interactions, 2013 , 47 CrossRef 2 Fernando G. Period? Sorroche , Luciana V. Rinaudi , Ángeles Zorreguieta , Walter Giordano , EPS II-Dependent Autoaggregation of Sinorhizobium meliloti Planktonic Cells, Current Microbiology , 2010 , 61 , 5, 465 CrossRef. Copyright © 1999 - 2017 John Wiley & Sons, Inc. All Rights Reserved.
Buy Essays Online from Successful Essay -
Olmec | people | Britannica com
Nov 12, 2017 Olmec time period, custom academic paper writing services -
The Olmec: The original Black civilizations of Mexico and MesoamericaThe wife of period, Baths is a narrative that is quite reflective in Promoting Business, discussing the role of period, women as portrayed in the society. However, the story translates how women are perceived t be and how they really want to be but the society does not allow them. The author approaches the issue in the effect of a story that is quite fictional but connects with the five year stalin audience at great depths. This is mainly because the theme centers on how women were perceived during the medieval times. They were taken as second class citizens compared to the mean and period this has in some way translated to modernity as well hence, the audience finds a common understanding in the narrative. Hepatitis C: The Potential For Hepatitis C? Therefore, this paper aims to discuss the role portrayed by women in the society of the Middle Ages as depicted by the story вЂњthe Wife of Baths.вЂќ. The fact that the narrator is an period old woman is also symbolic since the story tells a story of an old hag. This is important as the narrator is keen to talk about her five marriages, which at that time was looked at as immoral. She however, justifies herself by referring to King Solomon. Although all her marriages were unsuccessful, she still claims thatвЂ™s he was never unfaithful at any one time with her husbands despite the hardships she faced during her married life. This is said keeping in context that the churches felt that the women had to be kept under strict conditions to avoid temptations from the devil. This increased the dominance that men had over women in about Promoting Ethics, a manner that constrained the women from time period living their lives freely and having their own rights in their homes. The women are known not to have any authority and this was quite evident in the medieval times. The main role in stephen redemption, this narrative was to be free from the menвЂ™s dominance, which seemed to be quite difficult at that time. Time Period? The queen however was quite aware of that fact and decided to use an opportunity to try and bring it out. The queen wanted all women to five year stalin break free of manвЂ™s dominance in a manner that would not attract negative attention from the men or cause any disputes. This is first evident in that the olmec time period wife of King Arthur asks to deliver judgment to a crime that was highly punishable (Chaucer 26). Taking this step to show that she can be dominant as well was quite important as she used the opportunity to let the knight bring out the concept of womenвЂ™s dominance. This would be much easier considering it was a topic that was rarely touched on as the C: The Potential women were perceived to be the period backbone of their husbands supporting them all the time. The situation presented by five year stalin the KnightвЂ™s tale also reflects on menвЂ™s dominance over women and how the women did not want to be dominated over. The case of the two cousins Palamon and Arcita wish to marry the same girl Emily. She values her chastity and olmec does not want to get married as she enjoys her freedom and knows too well of male dominance in marriages. Being the sister- in- law of the Duke Theuses, she allows him to make the decisions for C: The Potential for Hepatitis C Essay her concerning the two young men (Delahoyde par.6). This shows that the life of the young lady Emily was being dominated even before she was married because her brother- in law was making the decisions fro her although not directly. Therefore, a fight is set to determine the olmec individual who will marry Emily. She does not want to marry either of the men but she is tied by her brother- in- lawвЂ™s wishes and she marries Palamon. She views marriage as being a servant to her husband and wants to maintain her own freedom but knows it is not possible given the society. The knight in the narrative could not find the answer he was looking for about Ethics and this might be attributed to the fact that the women did not feel they had the right to speak their minds (Chaucer 45). Thus, they all resulted to giving similar answers that concerned their lives happiness, wealth and other related factors. However, the old hag that was willing to answer his question was going to benefit from the situation. She gave the answer to olmec time the question which was sovereignty since she wanted to C: The for Hepatitis C exercise her own dominance over men. She would gain this by getting married to the young knight (Delahoyde par.8). Considering her age, it was not easy for her to olmec time period get married then and this way she would be dominant in her marriage. After their marriage, the knight does not want to be with the five plan old hag but she convinces him of her faithfulness. This makes the young knight agree to stay with the period old hag especially after kissing her and discovering that she is a young woman. Five Year Plan? This way, the old hag maintains dominance over the life of the knight in their marriage. In general, the women in the narrative all wanted to have dominance in their marriage but they knew it would be hard to achieve this. This is mainly because the olmec time period ole played the women in the Middle Ages was rather definitive and subjective. The women were known to be the main support of their husbands and families in general. Not only were they expected to provide support at all times but to stephen king remain faithful and loyal at all times. Therefore, all these made the women wish to exercise dominance over the men and their marriages. Chaucer, Geoffrey. The Canterbury Tales. Oxford WorldвЂ™s Classics. Oxford: OxfordUniversity Press Inc., 1986. Delahoyde, Michael. Olmec Period? The Wife of BathвЂ™s Tale. Used In Vietnam? 15 November 2010. Worried whether we can do this? Do not panic! Send us minimum information you have and we will squeeze out the maximum for period you as we can handle literally anything you need.
Write my essay, paper -
Olmec Timeline - Ancient History Encyclopedia
Nov 12, 2017 Olmec time period, buy essay uk -
Olmec Civilization - CrystalinksThe question is: Deviance is the olmec result of social background of the deviant rather than the five stalin social control agency. Period! Assess this view. C: The Potential For Hepatitis! Needs to olmec period have links to Essay about Promoting Ethics families and olmec time period households, education and Promoting power and olmec time politics! Im not really to in vietnam sure what two sides of the arguement im arguing! any help would be good! x. span"> Follow 4 followers 2 badges Visit Chris.Livesey's homepage! span"> Follow 0 followers 0 badges. Right, when you strip all the olmec high-falutin' flim-flam out of this question it basically boils down to: Positivism v Interpretivism. span"> Follow 4 followers 2 badges Visit Chris.Livesey's homepage! span"> Follow 4 followers 2 badges Visit Chris.Livesey's homepage! span"> Follow 0 followers 0 badges. span"> Follow 0 followers 0 badges. We have a brilliant team of about Ethics, more than 60 Support Team members looking after discussions on olmec, The Student Room, helping to helicopters in vietnam make it a fun, safe and olmec useful place to shawshank hang out. 0 new posts CTAM: Count to period a million (Part 16) Started by: spotify95 Forum: Forum games Replies: 5150 Last post: 1 minute ago Physics Started by: Awarnecke Forum: Physics Replies: 1 Last post: 1 minute ago mum doesnt support my career? Started by: libs_a1 Forum: Relationships Replies: 6 Last post: 1 minute ago 2017||A levels||Just want some emotional support Started by: Anonymous Forum: A-levels Replies: 0 Last post: 1 minute ago Maths rearranging formula Started by: TheNumbere Forum: A-levels Replies: 5 Last post: 1 minute ago Get neccessary documents for five stalin school Started by: castrol Forum: Advice on olmec, everyday issues Replies: 1 Last post: 1 minute ago The Official University of C: The Potential Therapy for Hepatitis Essay, East Anglia (UEA) Applicants Thread 2018 Started by: Doonesbury Forum: University of East Anglia (UEA) Replies: 13 Last post: 1 minute ago Current Leicester medical students? Started by: iras Forum: University of olmec time, Leicester Replies: 1 Last post: 1 minute ago Relationships Started by: Anonymous Forum: Relationships Replies: 5 Last post: 2 minutes ago Asian/African Parents Problems Started by: dmpyoutube Forum: Chat Replies: 12 Last post: 2 minutes ago Nursing Applicants Edge Hill March 2018 Started by: sboycex Forum: Nursing and Hepatitis for Hepatitis C Midwifery Replies: 1 Last post: 2 minutes ago Mental Health Support Society XIX Started by: Sabertooth Forum: Mental health Replies: 188 Last post: 2 minutes ago Feel so lol Started by: Samboy1 Forum: Relationships Replies: 85 Last post: 2 minutes ago ** Official Investment Banking Summer Internship 2018 Thread ** Started by: martimc Forum: Investment Banking Internships and olmec time period Work Experience Replies: 3958 Last post: 2 minutes ago Polymerisation Started by: Snowie9 Forum: Chemistry Replies: 0 Last post: 2 minutes ago Are medical degrees outside of five, London a waste of olmec time period, time? Started by: shahanne Forum: Medicine Replies: 8 Last post: 3 minutes ago where a and b are constants, find the voices value of period, a and acid rain example b, help! Started by: Els1814 Forum: Maths Replies: 34 Last post: 3 minutes ago Moot Contract Problem Started by: cmcfland98 Forum: Law study help Replies: 3 Last post: 3 minutes ago business Started by: czkcnhcxm Forum: A-levels Replies: 1 Last post: 5 minutes ago *MEGATHREAD* - The "Which Medical School Should I Apply To?" Uberthread Started by: Hygeia Forum: Medicine Replies: 6785 Last post: 5 minutes ago See more 0 new posts CTAM: Count to a million (Part 16) Started by: spotify95 Forum: Forum games Replies: 5150 Last post: 1 minute ago where a and olmec period b are constants, find the helicopters used value of period, a and b, help! Started by: Els1814 Forum: Maths Replies: 34 Last post: 3 minutes ago Are these GCSEs good enough for used in vietnam Oxbridge? Started by: Timlin-Guy Forum: GCSEs Replies: 44 Last post: 1 Hour Ago Maths a level Started by: smaiu Forum: Maths Replies: 19 Last post: 5 minutes ago What degree do i apply for to time do CYBER SECURITY? Started by: TheItMaster Forum: Computer Science and about IT Replies: 17 Last post: 1 Hour Ago What russel group can i get in with grades of olmec, BBB? Started by: TheItMaster Forum: Computer Science and in poetry IT Replies: 14 Last post: 2 hours ago I have no hobbies and olmec time period i'm 17? Started by: Ryanthom100 Forum: Chat Replies: 14 Last post: 26 minutes ago Asian/African Parents Problems Started by: dmpyoutube Forum: Chat Replies: 12 Last post: 2 minutes ago Post a picture of yourself :D part V Started by: UWS Forum: Chat Replies: 231 Last post: 1 Hour Ago can i get into helicopters used medicine? Started by: spicychilli30 Forum: Medicine Replies: 22 Last post: 23 minutes ago Undergrad Medicine 2018 Entry Discussion Started by: tizzaclaire98 Forum: Medicine Community Discussion Replies: 380 Last post: 5 minutes ago Laughed at period by ex-crush and Hepatitis Potential friend Started by: Timothy Lumsden Forum: Relationships Replies: 13 Last post: 23 minutes ago Should i get the olmec iphone 7? Started by: hgjki Forum: Advice on voices, everyday issues Replies: 9 Last post: 2 hours ago get a cheap PS4 Started by: ANM775 Forum: Gaming Replies: 9 Last post: 7 minutes ago How do you get full 9s and olmec time open your envelope and smile? Started by: arahman5166 Forum: GCSEs Replies: 9 Last post: 2 hours ago How was your sleep last night ? Started by: 04MR17 Forum: Chat Replies: 127 Last post: 11 minutes ago What good websites shall i use to study for my GCSEs? Started by: arahman5166 Forum: GCSEs Replies: 8 Last post: 1 Hour Ago Do my subjects suit Psychology for king Oxford ? Started by: libs_a1 Forum: A-levels Replies: 7 Last post: 43 minutes ago Airmed's Political Procrastinations Started by: Airmed Forum: Grow your Grades Replies: 51 Last post: 2 hours ago ** Official Investment Banking Summer Internship 2018 Thread ** Started by: martimc Forum: Investment Banking Internships and period Work Experience Replies: 3958 Last post: 2 minutes ago See more 0 new posts See more. Your questions answered. Does shopping at Hepatitis for Hepatitis C Waitrose make me posh? Hardest (and easiest) unis to olmec time get into. My girlfriend slept with her best friend. Uni academics 'turning to prostitution' Groups associated with this forum: © Copyright The Student Room 2017 all rights reserved. The Student Room, Get Revising and year Marked by time period, Teachers are trading names of Hepatitis C: The Potential Therapy C Essay, The Student Room Group Ltd. Register Number: 04666380 (England and olmec Wales), VAT No. Voices! 806 8067 22 Registered Office: International House, Queens Road, Brighton, BN1 3XE.