Its termini contain the inverted repeat sequence 5′-CCTGC … GCAGG-3′, and its 5′-ends are covalently capped with protein (Overhage et al., 2005; Parschat et al., 2007). Our previous sequence analysis of pAL1 and predictions of possible secondary structures formed by potential telomeric 3′-overhangs indicated significant differences of the ‘left’ and ‘right’ terminus of pAL1, raising the question of whether each terminus of pAL1 is recognized, or even capped, by a specific protein (Parschat et al., INNO-406 2007). Rhodococcal plasmids pHG201 and pHG205 are other examples of actinomycete

linear plasmids that do not show striking homology between their ‘left’ and ‘right’ telomere sequences (Kalkus et al., 1998), but their TPs have not been described. In contrast, the

ends of Streptomyces this website linear replicons usually contain well-conserved terminal palindromic sequences (Zhang et al., 2006). The gene product of pAL1.102 is the only protein exhibiting a weak similarity to known (Streptomyces) TPs; however, due to the low sequence similarity, its annotation as a ‘putative terminal protein’ was tentative (Parschat et al., 2007). As a first step toward characterizing the telomere complex of pAL1, we identified the protein attached to both termini of pAL1 and demonstrated its specific deoxynucleotidylation in vitro. The strains and plasmids used in this study are listed in Table 1. For isolation of total DNA, A. nitroguajacolicus Rü61a [pAL1] was grown in a mineral salts medium (Parschat et al., 2003) on 8 mM sodium benzoate at 30 °C. Arthrobacter nitroguajacolicus Rü61a [pAL1, pART2malE-ORF102 or pART2malE-ORF103] was cultivated in a mineral salts medium supplemented with 4 mM 4-hydroxyquinaldine SSR128129E and 140 μg mL−1 kanamycin. Cells were harvested by centrifugation at an OD600 nm of approximately 2.5. Escherichia coli DH5α clones containing derivatives of pMal-c2x or pART2 were grown in lysogeny broth (LB) (Sambrook & Russell, 2001) at 37 °C in the presence of 100 μg mL−1 ampicillin or 50 μg mL−1 kanamycin, respectively. For the synthesis of fusion

proteins of maltose-binding protein (MBP) and the protein encoded by pAL1.102 (termed pORF102), E. coli K12 ER2508 [pLysSRARE] harboring pMal-c2x-ORF102 was grown in LB with ampicillin (100 μg mL−1), chloramphenicol (34 μg mL−1), and auto induction solutions ‘5052’ and ‘M’ (Studier, 2005) at 30 °C. Cells were harvested by centrifugation at an OD600 nm of ∼5 and stored at −80 °C before use. Total DNA of A. nitroguajacolicus Rü61a [pAL1] was isolated according to Rainey et al. (1996). Plasmid DNA was isolated using the EZNA Plasmid Miniprep kit (Peqlab, Erlangen, Germany). Gel extraction of DNA fragments from agarose gels was performed with the Perfectprep gel cleanup kit (Eppendorf, Hamburg, Germany). For cloning purposes, DNA fragments were purified using the High Pure PCR Product Purification kit (Roche Diagnostics GmbH, Mannheim, Germany).

thuringiensis HD-1), hybridization experiments were performed using fragments of the DAPT in vitro three most abundant IS elements (IS231C, IS232A and ISBth166) of different families as probes (Fig. 1). There are no sites for Bst1107I and EcoRI within these elements. The result showed that

the most multitudinous IS231C had slightly different hybridization profiles between these two isolates, which indicated that it did not cause large-scale rearrangements. The observed variations in band patterns for IS232A and ISBth166 may result from homologous recombination between copies of these elements on chromosome or plasmids or may just be a result of their transposition to different locations. The increased number of identical copies within the YBT-1520 genome, numerous insertion events within functional genes and the identical positioning in different isolates suggest the recent expansion of IS231C. To explore the mobility of these three IS elements, we examined the changes in their Southern blot pattern during 30 repeated passages (bacterial generations) of YBT-1520 in vitro, which showed completely identical patterns, suggesting that these elements are relatively stable (data not shown). As two molecular markers of Btk were found in ISBth166 and PCR experiments indicated its existence in non-kurstaki

strains, the distribution of ISBth166 among B. thuringiensis NU7441 clinical trial serovars was further examined by Southern 17-DMAG (Alvespimycin) HCl blot analysis (Fig. 2). The restriction enzyme EcoRI was selected for B. thuringiensi genomic DNA digestions because

of the clear banding patterns. The resulting fragments were separated by agarose gel electrophoresis and probed with ISBth166 (see Materials and methods). The hybridization patterns showed that the five kurstaki strains hybridized strongly with the ISBth166 probe, while the other strains showed weak or null hybridization, suggesting that ISBth166 is widely distributed among kurstaki strains. The weak hybridization signal among some non-kurstaki strains may indicate the presence of ISs distantly related to ISBth166 or may reflect a lower copy number of the target sequence. The hybridization profiles of kurstaki strains showed a slight variation in the number and position of bands, while the strong hybridization signals may indicate the plasmid-borne elements that had more than one copy. Strains HD-231 and HD-232 showed the same hybridization patterns. The hybridization pattern of YBT-1520 was most similar to that of HD-263, with only one additional band. This result yielded useful indications for further experimental work to reveal the possible correlation between Lepidopteran larvae toxicity and the presence of ISBth166-related elements and the evolutionary relationships among the ISBth166 variants. In this study, 68 intact copies of 14 distinct IS elements were identified in the B.

It was mailed to 2,605 households; 1,704 responses were received, yielding a 65.4% response rate. A small incentive (monetary value less than $5) was given if the survey was completed and Dapagliflozin manufacturer returned by August 2008. Youthstyles data were weighted to reflect age and sex of child, household size, household income, head of household age, and race/ethnicity of adult of the US population, as determined by the 2007 Census estimates taken from the Current Population Survey. A traveler to a nonindustrialized country (from now on referred

to as “traveler”) was defined as a respondent who reported traveling in the last 12 months to a destination other than the United States, Canada, Europe, Japan, Australia, or New Zealand. Risk-taking attitude was measured by using a four-item Brief Sensation-Seeking Scale (BSSS-4) derived from the BSSS.8 The four items of the BSSS-4 are designed to assess four previously identified factors that comprise the construct of sensation seeking: experience seeking, disinhibition, thrill and adventure seeking,

and boredom susceptibility. The four items (questions 8–11, Table 1) of the BSSS-4 were scored continuously (1–4), providing a total sensation-seeking score ranging from Talazoparib research buy 4 to 16. Descriptive statistics of frequencies and percentages were analyzed. Fisher’s exact test was used for categorical variables, while Wilcoxon rank-sum test was used for continuous variables. p Values ≤0.05 were considered significant. Bivariate and multivariate logistic regressions were done to calculate odds ratios and 95% confidence intervals for demographic characteristics, with the final multivariate model determined using backwards elimination at a 5% significance level for variable selection. Cronbach’s coefficient alpha Mephenoxalone was used to determine internal consistency reliability for the four subscale survey questions. All analyses were done by using SAS software (Version 9.2; SAS Institute, Cary, NC, USA).

Of the 1,704 respondents, 131 (7.7%) had traveled in the previous 12 months to a nonindustrialized country. The mean age of travelers was 14 years old, and 59% of those who traveled were female (Table 2). Females were more likely to travel than males (p = 0.01). Compared with other variables, travel was also more positively associated with increasing household income (p < 0.0001), marital status of parents (p = 0.007), and increasing household size (p = 0.03). The multivariate model showed that the only significant factors associated with travel were sex (p = 0.01) and household income (p < 0.0001) (Table 2). The regions most often visited were Mexico (44.3%), the Caribbean (42.4%), and Central/South America (12%). The majority traveled for vacation (81.0%), followed by visiting friends or relatives (21.7%) and research/student (5.8%). Nearly one fifth of youth travelers (18.0%) traveled without their parents (Table 3).

, 2000), cystathionine α-synthase (His-Cys, Ojha et al., 2000), cytochrome P450cam (Cys-H2O, Dawson et al., 1982) and NO synthase (Dawson et al., click here 1982; Tsai et al., 1996). In contrast, most cytochromes c participating in electron transfer have His/Met or His/His coordination (Wilks, 2002). The His/Cys coordination in heme c is known to be limited: the aforementioned

SoxAX, the 40 kDa triheme cytochrome PufC in the photosynthetic reaction center (Alric et al., 2004), and the 15 kDa DsrJ in sulfate respiration (Pires et al., 2006) have the axial coordination. NaxL and NaxS have no homology to these proteins in the primary structure and the physiological roles of these proteins seem to be distinctly different. Nevertheless, the His/Cys coordination in heme c might commonly contribute to the protein functions. One possibility of such a contribution is to create the very low redox potential of heme. The two hemes in a SoxA subunit of P. pantotrophus have low redox potentials: one is −432 mV and the other is lower than that (Reijerse

et al., 2007). The heme c in DsrJ is also reported to have a low redox potential. The relatively high σ-donor ability of thiolate ligand, Cys-, effectively stabilizes the ferric state of heme, and conceivable polar surroundings would make the heme–iron redox potential further lower. Taken together, NaxLS of the anammox bacterium strain KSU-1 appears to be a novel member of c-type heme proteins with His/Cys axial coordination and a low redox

potential. The low redox potential of NaxLS reminds Fulvestrant research buy us of its potential role as an electron transmitter in anammox bacteria, in which electrons with a very low redox potential are generated on oxidation of hydrazine catalyzed by HZO and/or HAO, or on ferredoxin oxidation–reduction that is supposed to occur in anammox processes in C. Kuenenia stuttgartiensis (Strous et al., 2006). The physiological role of the NaxLS protein has not been elucidated as yet and further investigation is required. Appendix S1. Procedure to determine the nucleotide 5-Fluoracil supplier sequence of metagenomic fragment containing genes for NaxLS. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Three pathogens, Campylobacter, Salmonella, and Shiga-toxin-producing Escherichia coli, are leading causes of bacterial gastroenteritis in the United States and worldwide. Although these three bacteria are typically considered food-borne pathogens, outbreaks have been reported due to contaminated drinking water and irrigation water. The aim of this research was to develop two types of PCR assays that could detect and quantify three pathogens, Campylobacter spp., E. coli O157:H7, and Salmonella spp., in watershed samples.

, 2000), cystathionine α-synthase (His-Cys, Ojha et al., 2000), cytochrome P450cam (Cys-H2O, Dawson et al., 1982) and NO synthase (Dawson et al., Gefitinib mouse 1982; Tsai et al., 1996). In contrast, most cytochromes c participating in electron transfer have His/Met or His/His coordination (Wilks, 2002). The His/Cys coordination in heme c is known to be limited: the aforementioned

SoxAX, the 40 kDa triheme cytochrome PufC in the photosynthetic reaction center (Alric et al., 2004), and the 15 kDa DsrJ in sulfate respiration (Pires et al., 2006) have the axial coordination. NaxL and NaxS have no homology to these proteins in the primary structure and the physiological roles of these proteins seem to be distinctly different. Nevertheless, the His/Cys coordination in heme c might commonly contribute to the protein functions. One possibility of such a contribution is to create the very low redox potential of heme. The two hemes in a SoxA subunit of P. pantotrophus have low redox potentials: one is −432 mV and the other is lower than that (Reijerse

et al., 2007). The heme c in DsrJ is also reported to have a low redox potential. The relatively high σ-donor ability of thiolate ligand, Cys-, effectively stabilizes the ferric state of heme, and conceivable polar surroundings would make the heme–iron redox potential further lower. Taken together, NaxLS of the anammox bacterium strain KSU-1 appears to be a novel member of c-type heme proteins with His/Cys axial coordination and a low redox

potential. The low redox potential of NaxLS reminds Tacrolimus in vivo us of its potential role as an electron transmitter in anammox bacteria, in which electrons with a very low redox potential are generated on oxidation of hydrazine catalyzed by HZO and/or HAO, or on ferredoxin oxidation–reduction that is supposed to occur in anammox processes in C. Kuenenia stuttgartiensis (Strous et al., 2006). The physiological role of the NaxLS protein has not been elucidated as yet and further investigation is required. Appendix S1. Procedure to determine the nucleotide Teicoplanin sequence of metagenomic fragment containing genes for NaxLS. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Three pathogens, Campylobacter, Salmonella, and Shiga-toxin-producing Escherichia coli, are leading causes of bacterial gastroenteritis in the United States and worldwide. Although these three bacteria are typically considered food-borne pathogens, outbreaks have been reported due to contaminated drinking water and irrigation water. The aim of this research was to develop two types of PCR assays that could detect and quantify three pathogens, Campylobacter spp., E. coli O157:H7, and Salmonella spp., in watershed samples.

These

findings suggest that: (i) Reelin regulates the somatosensory barrel cortex differently than other neocortical areas, (ii) most Dab1 medial septum/diagonal band neurons are probably GABAergic projection neurons, and (iii) positioning errors in adult mutant Dab1-labeled neurons vary from subtle to extensive. “
“Department of Pediatric Hematology/Oncology, University Children’s Hospital Bonn, Bonn, Germany Grünenthal GmbH, Aachen, Germany Centre for Cognitive Neuroimaging, Institute of Neuroscience and Psychology, University of Glasgow, Glasgow, UK Chicken acidic leucine-rich EGF-like domain-containing brain protein (CALEB), also known as chondroitin sulfate proteoglycan (CSPG)5 or neuroglycan C, is a neural chondroitin sulfate-containing and epidermal growth factor (EGF)-domain-containing transmembrane protein that is implicated Forskolin mw in synaptic maturation. Here, we studied the role of CALEB within the developing cerebellum. Adult CALEB-deficient mice displayed impaired motor coordination in Rota-Rod experiments.

Analysis of the neuronal connectivity of Purkinje cells by patch-clamp recordings demonstrated impairments of presynaptic maturation of inhibitory synapses. GABAergic synapses on Purkinje cells revealed decreased evoked amplitudes, altered paired-pulse facilitation and reduced depression after repetitive stimulation at early postnatal but not at mature stages. Furthermore, the elimination of supernumerary climbing fiber synapses on Purkinje cells was found to occur at earlier Epacadostat in vitro developmental stages in the absence of CALEB. For example, at postnatal day 8 in wild-type mice, 54% of Purkinje cells had three or more climbing fiber synapses in contrast to C225 mutants where this number was decreased to less than 25%. The basic properties of the climbing fiber Purkinje cell synapse remained unaffected. Using Sholl analysis of dye-injected Purkinje cells we revealed that

the branching pattern of the dendritic tree of Purkinje cells was not impaired in CALEB-deficient mice. The alterations observed by patch-clamp recordings correlated with a specific pattern and timing of expression of CALEB in Purkinje cells, i.e. it is dynamically regulated during development from a high chondroitin sulfate-containing form to a non-chondroitin sulfate-containing form. Thus, our results demonstrated an involvement of CALEB in the presynaptic differentiation of cerebellar GABAergic synapses and revealed a new role for CALEB in synapse elimination in Purkinje cells. “
“During post-weaning development, a marked increase in peer–peer interactions is observed in mammals, including humans, which is signified by the abundance of social play behaviour. Social play is highly rewarding, and known to be modulated through monoaminergic neurotransmission.

Amino acid sequences for the homologous proteins were obtained from NCBI and TIGR databases [National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov) and the Institute of Genomic Research (http://www.tigr.org)]. Multiple sequence alignments were generated using the clustalw web-based program with default parameters [European Bioinformatics institute (http://www.ebi.ac.uk/clustalw)]. A model of putative NarP protein was made based on the crystal structure of E. coli NarL (Baikalov et al., 1996). After the putative M. haemolytica

NarP and E. coli NarL was aligned, the amino acids of the E. coli NarL was substituted with OSI-744 mouse the corresponding one of the M. haemolytica NarP using deepview/swiss-pdbviewer (http://www.expasy.org/spdbv/; version 3.7). After the model was optimized with the same software, it was visualized using macpymol Ribociclib clinical trial (DeLano Scientific LLC; http://delanoscientific.com/; version 0.98). The construction of narP mutants was carried out as described in McKerral

& Lo (2002) and the narP mutants were selected according to the protocol of Fedorova & Highlander (1997b) (see Supporting Information). The growth characteristics of MhΔNarP7 in comparison with the parent SH1217 and their response to nitrate were examined. An overnight culture of SH1217 or MhΔNarP7 was diluted 1/100 into BHIB, with or without NaNO3 supplementation. Five-milliliter aliquots of this culture were added to 15 test tubes and grown semi-anaerobically at 37 °C. The OD600 nm of the cultures were determined Cediranib (AZD2171) over 8 h at 2-h intervals, taking measurements from three test tubes at each interval. The OD600 nm values of different strains/culturing conditions were compared using an unpaired, two-tailed t-test (P<0.005). SH1217 and MhΔNarP7 were grown in 5 mL BHIB with or without NaNO3 supplement, semi-anaerobically at 37 °C. The cells were harvested at OD600 nm of 0.5. Total protein preparations were prepared by adding equal volume of 2 × sodium dodecyl sulfate polyacrylamide gel electrophoresis

(SDS-PAGE) loading buffer with the cell suspension and examined by SDS-PAGE and Western immunoblot according to Lo & Mellors (1996). After SDS-PAGE, the proteins were stained with Commassie brilliant blue. For Western immunoblot, the proteins were transferred to a nitrocellulose membrane as described (Lo et al., 1991), and blocked by immersion in a 3% gelatin solution in Tris-HCl-buffered saline containing 0.05% Tween 20 (TTBS). The Lkt neutralizing monoclonal antibody 601 (Gentry & Srikumaran, 1991) was used at a dilution of 1/2000 in antibody solution (1% gelatin in TTBS). The secondary antibody goat anti-mouse alkaline phosphates conjugate (Jackson Laboratories) was used at a dilution of 1/5000 in antibody solution. The membranes were developed using 5-bromo-4-chloro-3-indoyl-phosphate and nitroblue tetrazolium.

Amino acid sequences for the homologous proteins were obtained from NCBI and TIGR databases [National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov) and the Institute of Genomic Research (http://www.tigr.org)]. Multiple sequence alignments were generated using the clustalw web-based program with default parameters [European Bioinformatics institute (http://www.ebi.ac.uk/clustalw)]. A model of putative NarP protein was made based on the crystal structure of E. coli NarL (Baikalov et al., 1996). After the putative M. haemolytica

NarP and E. coli NarL was aligned, the amino acids of the E. coli NarL was substituted with EPZ015666 the corresponding one of the M. haemolytica NarP using deepview/swiss-pdbviewer (http://www.expasy.org/spdbv/; version 3.7). After the model was optimized with the same software, it was visualized using macpymol Ruxolitinib purchase (DeLano Scientific LLC; http://delanoscientific.com/; version 0.98). The construction of narP mutants was carried out as described in McKerral

& Lo (2002) and the narP mutants were selected according to the protocol of Fedorova & Highlander (1997b) (see Supporting Information). The growth characteristics of MhΔNarP7 in comparison with the parent SH1217 and their response to nitrate were examined. An overnight culture of SH1217 or MhΔNarP7 was diluted 1/100 into BHIB, with or without NaNO3 supplementation. Five-milliliter aliquots of this culture were added to 15 test tubes and grown semi-anaerobically at 37 °C. The OD600 nm of the cultures were determined aminophylline over 8 h at 2-h intervals, taking measurements from three test tubes at each interval. The OD600 nm values of different strains/culturing conditions were compared using an unpaired, two-tailed t-test (P<0.005). SH1217 and MhΔNarP7 were grown in 5 mL BHIB with or without NaNO3 supplement, semi-anaerobically at 37 °C. The cells were harvested at OD600 nm of 0.5. Total protein preparations were prepared by adding equal volume of 2 × sodium dodecyl sulfate polyacrylamide gel electrophoresis

(SDS-PAGE) loading buffer with the cell suspension and examined by SDS-PAGE and Western immunoblot according to Lo & Mellors (1996). After SDS-PAGE, the proteins were stained with Commassie brilliant blue. For Western immunoblot, the proteins were transferred to a nitrocellulose membrane as described (Lo et al., 1991), and blocked by immersion in a 3% gelatin solution in Tris-HCl-buffered saline containing 0.05% Tween 20 (TTBS). The Lkt neutralizing monoclonal antibody 601 (Gentry & Srikumaran, 1991) was used at a dilution of 1/2000 in antibody solution (1% gelatin in TTBS). The secondary antibody goat anti-mouse alkaline phosphates conjugate (Jackson Laboratories) was used at a dilution of 1/5000 in antibody solution. The membranes were developed using 5-bromo-4-chloro-3-indoyl-phosphate and nitroblue tetrazolium.

The origin of index cases was highly consistent with population migration routes and countries most frequently visited by French tourists as shown by a nationwide study.[6] This fact justifies screening and presumptive isolation with contact precautions of patients transferred from or previously hospitalized abroad.[10] Although returned travelers who have neither been ill nor hospitalized during travel can acquire resistant bacteria, at present, the risk does

not seem high enough to justify routinely screening all hospitalized patients with history of recent travel. If the number of CPE events

increased between 2004 and 2011, the number of outbreaks EPZ015666 remained low, eg, only three outbreaks occurred in 2011, contrasting with 40 reported events during the same period. This fact emphasizes the efficacy of the specific control measures of the AP-HP “emergent MDR” program[7, 9] and specially screening and isolating patients transferred from foreign hospitals. The AP-HP program has been subsequently extended at the national level by health authorities.[10] Importantly, intensive care units are the first wards concerned by repatriation of CPE carriers patients in our study and physicians must be

alerted of this risk. In conclusion, proactive strategy Crizotinib chemical structure to control spread of antibiotic resistance such as CPE should include systematic screening and isolation of patients transferred from or previously hospitalized abroad. The authors state that they have no conflicts of interest to declare. Antoine Andremont, Frédéric Barbut, Edouard Bingen, Christine Carbohydrate Bonnal, Emmanuelle Cambau, Anne Carbonne, Anne Casetta, Jacques Chemardin, Jean-Winoc Decousser, Catherine Doit, Florence Doucet-Populaire, Laurence Drieux-Rouzet, Florence Espinasse, Nicolas Fortineau, Jean-Louis Gaillard, Jean-Michel Guérin, Laurent Gutmann, Béate Heym, Guillaume Kac, Christine Lawrence, Patrick Legrand, Jean-Christophe Lucet, Simone Nerome, Marie-Hélène Nicolas-Chanoine, Patrice Nordmann, Jean-Claude Petit, Bertrand Picard, Claire Poyart, Laurent Raskine, Jérôme Robert, Martine Rouveau, Delphine Seytre, and Isabelle Simon. “
“Background. Increasing air travel has resulted in a significant increase in aeromedical evacuation (AE) over the past decade. However, there are limited epidemiological data available on the diagnosis, costs, and transport characteristics of AE cases. Methods.

Other physiological characteristics of the isolate were tested with API 20NE and API 50CH test strips (bioMérieux). API 20NE and API 50CH tests results

were observed over a period of 7 days at 25 °C. Antibiotic sensitivity was tested by spreading a bacterial suspension on R2A and applying discs impregnated with the following antibiotics (concentration per disc): selleckchem ampicillin (10 μg), amikacin (30 μg), ceftriaxone (30 μg), clindamycin (2 μg), gentamicin (30 μg), kanamycin (30 μg), neomycin (30 μg), penicillin (10 μg), streptomycin (10 μg), tetracycline (30 μg) and vancomycin (30 mg). Isoprenoid quinones of strain DR-f4T were analyzed with freeze-dried cells previously grown in R2A for 3 days according to the method of Collins & Jones (1981) and Komagata & Suzuki (1987). The quinone was purified via preparative thin-layer chromatography (silica gel F254; Merck) and was identified using an HPLC (Hitachi L-5000) equipped with a reverse-phase column (YMC pack ODS-AM; YMC Co.). For fatty acid methyl esters (FAMEs) analysis, strain DR-f4T was cultured on R2A (pH 6.0) at 20 °C for 3 days, which are the same culture conditions as those used for FAMEs analysis of the closest type strain, M. lappiensis ANJL12T (Männistöet al., 2010). Selleck CYC202 FAMEs were extracted according to the standard protocol of the microbial identification system (MIDI;

Sasser, 1990), separated by a gas chromatograph (HP 6890N; Agilent) and identified using the sherlock software package (MIDI). Genomic DNA of strain DR-f4T and E. coli KCTC 2441T was extracted according to the method described PAK6 by Sambrook & Russell (2001). The G+C content of the isolate was determined using the method described by Mesbah et al. (1989). Briefly, genomic DNAs were hydrolyzed and dephosphorylated with nuclease P1 and with alkaline phosphatase, respectively, and then the mixtures of nucleosides were analyzed by HPLC for G+C mol%. The 16S rRNA gene was amplified by PCR with the universal primers 27F and 1492R (Lane, 1991). After

purification of the PCR product, the sequencing reaction of the 16S rRNA gene was performed at SolGent Co., Korea, using an ABI prism Bigdye terminator cycle sequencing ready reaction kit V.3.1 and an ABI 3730XL capillary DNA Sequencer (Applied Biosystems). The sequence of the 16S rRNA gene was assembled using vector nti software (Invitrogen). The sequence of strain DR-f4T was compared with available 16S rRNA gene sequences from the GenBank using the blast program (http://www.ncbi.nlm.nih.gov/blast/) and the EzTaxon server (http://www.eztaxon.org/; Chun et al., 2007). The 16S rRNA gene sequence of strain DR-f4T was aligned with those of representative members of selected taxa belonging to the family Sphingobacteriaceae using the clustal_x software (Thompson et al., 1997), and this alignment was edited manually.