These differences may reflect profound differences in the regulat

These differences may reflect profound differences in the regulation of some proteases between human and porcine thyrocytes because the expression of DPP IV and APN is linked to transformation of human thyrocytes. Thyrocytes from animals play an important role in the study of physiological processes because inter-individual variations in animals usually are lower than in humans. Inter-individual variations in protein expression, iodide uptake, proliferation and other physiological reactions are more pronounced in normal DAPT chemical structure human thyroid tissue

samples and isolated human thyrocytes than in porcine ones [31]. Causes of different physiological reactions among individuals include genetic factors and environmental factors (dietary iodine, smoking, infections, etc.). Due to these limitations, porcine, bovine, ovine and canine thyrocytes are common substitutes for normal human cells because only animal thyrocyte lines able to form follicles and synthesize

thyroid hormones are available [1, 32]. Despite general similarities in morphology, synthesis of thyroid hormone and the reaction to TSH, several differences between the species, including molecular differences in proteins, in expression and in reaction to growth factors, have been identified [2, 33–36]. In our study, lysosomal protease activity of DPP II was strongly expressed in thyrocytes of all species. This lack of interspecies PRIMA-1MET manufacturer Thalidomide differences was also reported in another study on the expression pattern of the lysosomal proteases cathepsin B and elastase in the placenta of mice, rats, guinea pigs and marmosets [37]. In contrast, we saw expression of DPP IV and APN only in porcine thyrocytes but not in thyrocytes from other species. In human thyroid glands, consistent with previous studies, thyrocytes lacked both enzyme activities

and only endothelial cells showed reactivity for DPP IV [38]. Pronounced interspecies variations in the expression of the membrane-associated proteases were also reported by Gossrau and Graf, who investigated cellular expression of γ-glutamyltranspeptidase, aminopeptidase A, APN and DPP IV activities [37]. The observed differences in protease activities persisted in cultured porcine cells when cultured in the presence of TSH. As the membrane-associated proteases DPP IV and APN localize to the apical NVP-BGJ398 research buy membrane, they are only expressed when follicles are formed. This indicates that, contrary to human thyrocytes, they are markers of differentiation, not de-differentiation. Expression of APN in porcine thyrocytes has also been reported by Feracci et al. [27]. Because of these observed differences, porcine thyrocytes are not suitable models for studies on the regulation of membrane-protease in human thyrocytes. The determination of actual protease activity in this study, instead of merely detecting protein or mRNA, allows a direct assessment of relevant functional activity.

After the system was sonicated for 30 min, TEOS of 4 mL (10 mM) w

After the system was sonicated for 30 min, TEOS of 4 mL (10 mM) was added to the above system and the entire system was stirred

for 20 h. Next, 10 mL APTS was added to form a mixed solution and allowed to react at 80°C for 3 h. The resultant product was treated by high-speed centrifugal separation and washed with deionized water for several times, and then dried at 60°C for 3 h in a vacuum oven to obtain the sGNRs. Fabrication of sGNR/MWNT nanohybrid Covalent attachment of sGNRs to the MWNTs was performed using a modification of the standard EDC/NHS reaction [49, 50]. Carboxyl groups on the surface of MWNTs (5 mg) were activated by an EDC/NHS solution for 30 min. Following activation, 1 mg of sGNRs were added to form a mixed solution and allowed to react

at room temperature for 6 h, and then RGD peptides were added into Doramapimod clinical trial the mixed solution and continued to react at room temperature for 6 h. The resultant products were treated by high-speed centrifugal separation and washed with deionized water for three times, and then kept at 4°C for use. Characterization of sGNR/MWNT nanohybrid A JEOL JEM-2010 transmission electron microscope and a JEOL JEM-2100 F high-resolution transmission electron microscope (JEOL Ltd., Akishima, Tokyo, Japan) were used to confirm particle size and KPT330 observe the interface and the binding site of sGNRs and MWNTs. UV-vis spectra were measured at 20°C with a Shimadzu UV-2450 UV-visible spectrophotometer (Shimadzu

Corporation, Kyoto, Japan) equipped with a 10-mm quartz cell, where the light path length was 1 cm. The 200- to 1,000-nm wavelength region was scanned, since it includes the absorbance of the GNRs. The Fourier transform infrared (FTIR) spectra were recorded on a PerkinElmer Paragon-1000 FTIR spectrometer (PerkinElmer, Waltham, MA, USA). Zeta potential was measured with a Nicomp 380ZLS Zeta Potential/Particle Sizer (Nicomp, Santa Barbara, CA, USA). Effects of RGD-GNR-MWNT nanoprobes on cell viability Effects of RGD-GNR-MWNT nanoprobes on viability of MGC803 and GES-1 cells were analyzed using Cell Counting Kit-8 (CCK8) assay [23]. MGC803 and GES-1 cells were cultured in a 96-well microplate at Phospholipase D1 the concentration of 5,000 cells per well and incubated in a humidified 5% CO2 balanced air incubator at 37°C for 24 h. Except for control wells, the remaining wells were added into the medium with RGD-GNR-MWNT nanoprobes. Final concentrations were, respectively, 5, 10, 40, and 80 μg/mL, and then those cells were continuously cultured for 24 days. Then, the ODs were measured using the Thermo Multiskan MK3 ELISA plate reader (Thermo Fisher Scientific, Waltham, MA, USA) AZD8186 according to the protocol of the CCK8 assay kit, and the survival rate of cells was calculated.

metallireducens genome encodes 83 putative sensor histidine kinas

metallireducens genome encodes 83 putative sensor histidine kinases containing HATPase_c domains (Additional file 12: Table S7), of which 45 (54%) have orthologs among the 95 such CP673451 proteins of G. sulfurreducens. There are 94 proteins with response receiver (REC) SBE-��-CD manufacturer domains

in G. metallireducens (Additional file 12: Table S7), out of which 66 (70%) have orthologs among the 110 such proteins of G. sulfurreducens. Twenty-seven of the REC domain-containing proteins and another 101 genes and four pseudogenes (Additional file 12: Table S7) were predicted to be transcriptional regulators in G. metallireducens. There are 20 putative diguanylate cyclases containing GGDEF domains, of which 16 (80%) have orthologs among the 29 putative diguanylate cyclases LY411575 of G. sulfurreducens (Additional file 13: Table S8). Overall, the portion of the genome dedicated to signalling

and transcriptional regulation in G. metallireducens is slightly less than in G. sulfurreducens, but still considerable and significantly different in content. Several protein factors involved in chemotaxis-type signalling pathways are conserved between the two genomes: G. sulfurreducens and G. metallireducens each possess four or five CheA sensor kinases and ten CheY response receivers, almost all of which are orthologous pairs (Additional file 14: Table S9). In contrast, 17 of the 34 methyl-accepting chemotaxis proteins (MCPs) of G. sulfurreducens have no full-length matches in G. metallireducens (Additional file 14: Table S9). Due to apparent gene family expansion in G. sulfurreducens, its remaining 17 MCPs correspond to only 13 MCPs of G. metallireducens (Additional file 14: Table S9). The other five MCPs of G. metallireducens lack full-length matches in other Geobacteraceae (Additional file 14: Table

S9). Whereas G. sulfurreducens may use its closely related MCPs to fine-tune its chemotactic responses, G. metallireducens may accomplish response modulation by having twice as many MCP methyltransferases (CheR) and methylesterases (CheB) as G. sulfurreducens (Additional file 14: Oxalosuccinic acid Table S9). Integration host factors (IHF) and histone-like (HU) DNA-binding proteins are global regulators of gene expression composed of two homologous proteins that bend DNA in specific locations [117]. IHF/HU binding sites are favoured by some mobile genetic elements for insertion. The genome of G. metallireducens encodes orthologs of the single HU protein, both IHF beta proteins, and one of two IHF alpha proteins of G. sulfurreducens (Table 4). Another HU gene and two additional IHF alpha genes are present in G. metallireducens but not G. sulfurreducens (Table 4).

4) Prior to cell lysis for co-IP, washed cells (4 × 107 organism

4). Prior to cell lysis for co-IP, washed cells (4 × 107 organisms) from each culture condition were subjected to anti-BamA immunoblot analysis to verify the regulatable BamA phenotype. For co-IP experiments, cell pellets were solubilized and lysed by resuspension in 1× MRT67307 BugBuster Reagent (EMD Biosciences, Inc., Darmstadt, Germany; 2.5 mL per gram of wet cell weight). The solubilized cell solution was supplemented with 2 μL Lysonase Bioprocessing Reagent (EMD Biosciences,

Inc.) and 20 μL of protease inhibitor cocktail (Sigma Chemical Company, St. Louis, MO) per co-IP sample, and the mixture was subsequently rocked at room temperature Wnt/beta-catenin inhibitor (RT) for 20 min. Finally, the cell debris was pelleted at 15,000 × g for 15 min at 4°C, and the supernatant (containing

the cell lysate) was used for the co-IP experiments. Co-IPs were performed using the Sigma Protein G Immunoprecipitation Kit according to manufacturer’s instructions, with the following modifications: 1) the 1× and 0.1× IP Buffers were supplemented with 0.2% Triton X-100, and 2) prior to immunoprecipitation, the lysates were pre-cleared overnight to reduce background binding. After immunoprecipitation, bound proteins were eluted in 50 μL final sample buffer [62 mM Tris-HCl (pH 6.8), 10% v/v glycerol, 100 mM DTT, 2% SDS, 0.001% bromophenol blue], subjected to SDS-PAGE, and analyzed by silver stain according to the procedure of Morrissey [51], or by immunoblot, as described above. For protein identification, excised SDS-PAGE gel bands were submitted PKC inhibitor to the Molecular Biology-Proteomics Facility (University of Oklahoma HSC, Oklahoma City, OK) for tryptic digestion and HPLC-MS/MS analysis, followed by MASCOT database search for protein identification.

Triton X-114 (TX-114) phase partitioning To determine whether BB0324 and BB0028 have the amphipathic properties of typical lipid-modified proteins, B. burgdorferi strain B31-MI cells (2 × 108 organisms) were harvested and phase-partitioned as described previously [39, 52]. Proteinase K (PK) surface accessibility To determine whether BB0324 and BB0028 contain surface-exposed regions, PK experiments were performed as previously buy Baf-A1 described [39]. Briefly, spirochetes (2 × 108 organisms) were harvested at 4,000 × g, washed four times in 1× PBS (pH 7.4), and the washed cells were either mock-treated or PK-treated (400 μg/μl); Sigma Chemical Co.) for one hour at RT. After addition of PMSF (0.4 mM final concentration), samples were prepared for SDS-PAGE and immunoblot analysis, as described above. To verify that BB0324 and BB0028 were not resistant to PK activity, cell membranes were disrupted as previously described [53]. Cells (2 × 108 or 1 × 109) were pelleted at 10,000 × g, washed, and incubated for 10 m in 200 μl PK lysis buffer containing 50 mM Tris, 0.5% Triton X-100, 0.1%, β-mercaptoethanol, and 50 μg of lysozyme.

7 macrophage-like cells; CRL-2278; ATCC, Manassas, VA) were maint

7 macrophage-like cells; CRL-2278; ATCC, Manassas, VA) were maintained within a humidified VEGFR inhibitor environment at 37°C and under 5% CO2 in complete DMEM, (Thermo Scientific, Waltham, MA) containing penicillin (100 U; Gibco BRL, Grand Island, NY), streptomycin (0.1 mg/ml; Gibco BRL), L-glutamine (2 mM; Sigma, St. Louis, MO), and FBS (10%; JRH Biosciences, Lenexa, KS). MH-S cells (CRL-2019; ATCC) were maintained within a humidified environment at 37°C and under 5% CO2 in complete RPMI medium (Thermo Scientific) containing penicillin-streptomycin (100 U, Gibco BRL), L-glutamine (4 mM), and FBS (10%). JAWSII (CRL-11904; ATCC) were maintained within a humidified

environment at 37°C and under 5% CO2 in complete MEMα (Thermo Scientific) containing penicillin-streptomycin (100 U), L-glutamine (4 mM), and FBS (20%). RAD001 in vivo All tissue culture plasticware was purchased from Corning Incorporated (Corning, NY). Evaluation of B. anthracis spore germination in cell culture media Using 96 well plates, spores prepared from B. anthracis 7702 (1.0 × 108 spores/mL) were incubated at 37°C STA-9090 and under 5% CO2 in BHI (BD Biosciences, San Jose, CA), LB (0.1% tryptone, BD Biosciences; 0.05% yeast extract, BD Biosciences; 0.05% NaCl, Fisher Chemical, Fairlawn, NJ), PBS pH 7.2 (Mediatech, Manassas, VA), or germinating amino acids (10 mM L-alanine, 10 mM L-inosine, both from Sigma) in PBS pH 7.2. In other

studies, spores were incubated in 96 well plates (108 spores/mL) and at 37°C and under 5% CO2 in the following cell culture media without or with FBS (10%, unless otherwise indicated; Mediatech): DMEM (0.1, 0.5, 1, 5 or 10% FBS), RPMI-1640, MEMα modification (10 or 20% FBS), MEM (Mediatech), AMEM (Gibco), EMEM

(Mediatech), BME (Sigma), CIM (Gibco), Ham’s F-12 (Mediatech), McCoy’s 5A (M5A, ATCC), or DMEM with 10% FBS and 10 mM D-alanine (Sigma) and D-histidine (Sigma). In some assays, FBS obtained from Mediatech was substituted with FBS purchased from Invitrogen or Sigma. As described previously [39], spore germination was evaluated by measuring loss in spore refractility or loss of heat resistance, while outgrowth was monitored by monitoring the elongation of bacilli using a Delta Vision RT microscope (Applied Precision; Issaquah, WA), outfitted with an Olympus Plan Apo 100 × oil objective. DIC images were Farnesyltransferase collected using a Photometrics CoolSnap HQ camera; (Photometrics, Tucson; AZ), and processed using SoftWoRX Explorer Suite (version 3.5.1, Applied Precision Inc). Pre-conditioning of cell culture media To pre-condition cell culture medium, monolayers of RAW264.7 or MH-S cells in 24-well plates (80 to 95% confluency) were washed three times with Hanks’ balanced salt solution (HBSS) and then incubated in DMEM (for RAW264.7 cells) or RPMI-1640 (for MH-S cells) without FBS and penicillin-streptomycin in a humidified environment at 37°C and under 5% CO2.

Instead, they both have MFS-type nitrate/nitrite trans

Instead, they both have MFS-type nitrate/nitrite transporters (see above). Sco has about 4 times as many ABC amine transport proteins as does Mxa. These two organisms

have similar numbers of ABC iron uptake proteins (11 and 8, respectively). ABC uptake systems for inorganic cations are rare in both bacteria. Vitamin transporters are also scarce. ABC-type export systems are less numerous than uptake systems in both organisms. However, some families are well represented in one or the other organism. Both have at least one putative LPS precursor export system (Family 103), several lipid exporters (Family 106), and several I-BET-762 in vitro lipoprotein exporters (Family 125) (Table 10). ABC-type drug exporters are prevalent but with striking differences between the two organisms. Sco has ten DrugE1 export proteins (Family 105) while Mxa has only one. Both have a single DrugE2 exporter (Family 117), but while Sco has only one DrugE3 export protein (Family 119), Mxa has six. Most strikingly, while Sco has only one macrolid export protein (Family 122), Mxa has 16. They both have MDR pumps belonging to other ABC export families, including eukaryotic-type systems. In Mxa, two of these belong to the MDR Family (Family 201), while in Sco, 1 belongs to the EPP Family

(Family 204). Protein and peptide exporters can also be found, but no family predominates in either organism, and representation of one family in one of these selleck products bacteria does not correlate with representation in the other (Table 10). It seems clear that these two organisms 4-Aminobutyrate aminotransferase have solved the problems of macromolecular and drug export using very different transport systems and mechanisms. This fact probably reflects the independent evolution of the two sporulating organisms’ lifestyles, as well as the production and secretion of different types of molecules. Thus, in spite of their striking selleck chemical physiological similarities (see Discussion), Sco

and Mxa have used very different types of transport systems to satisfy their metabolic and developmental needs. Table 10 ABC export porters in Sco and Mxa TC # Family name Known substrate range ABC Type Sco Mxa 3.A.1.103 Lipopolysaccharide Exporter (LPSE) LPS 2 2 1 3.A.1.105 Drug Exporter-1 (DrugE1) Drugs 2 10 1 3.A.1.106 Lipid Exporter (LipidE) PL, LPS, Lipid A, Drugs, Peptides 1 6 3 3.A.1.107 Putative Heme Exporter (HemeE) Heme, Cytochrome c 2   1 3.A.1.109 Protein-1 Exporter (Prot1E) Proteins 1   1 3.A.1.110 Protein-2 Exporter (Prot2E) Proteins 1   1 3.A.1.111 Peptide-1 Exporter (Pep1E) Bacteriocin, Peptides 1 2 1 3.A.1.112 Peptide-2 Exporter (Pep2E) Other Peptides 1 1   3.A.1.115 Na+ Exporter (NatE) Sodium 2   1 3.A.1.117 Drug Exporter-2 (DrugE2) Drugs, Lipids, Dyes 1 1   3.A.1.119 Drug/Siderophore Exporter-3 (DrugE3) Drugs, Siderophores 1 6   3.A.1.122 Macrolide Exporter (MacB) Macrolides, Heme 3 1 16 3.A.1.123 Peptide-4 Exporter (Pep4E) Drugs, Peptides 1 1   3.A.1.

95) when compared to incubation without plasma (Figure 3), sugges

95) when compared to incubation A-769662 cell line without plasma (Figure 3), suggesting that the presence

of non-specific IgG does not alter the ability of hRS7 to mediate ADCC in Trop-2 expressing carcinosarcoma cells. Figure 3 Representative cytotoxicity experiments against the OMMT-ARK-2 cell line. Cytotoxicity in the presence of human plasma diluted 1:2 (with or without heat-inactivation) with effector cells and either hRS7 or rituximab control antibody in 5 h 51Cr-release assays. Addition of untreated plasma (diluted 1:2) to PBL in the presence of hRS7 significantly increased the ADCC achieved in the presence of hRS7 and PBL against OMMT-ARK-2 (P = 0.002). Addition of physiological concentrations of IgG (i.e. heat-inactivated plasma diluted 1:2) to PBL in the presence of hRS7 did not significantly alter the degree of ADCC achieved against OMMT-ARK-2 in the presence of hRS7 and PBL SAHA HDAC ic50 (P = 0.95). Discussion In this study, we have investigated Trop-2 expression Selleck CYC202 and localization by immunohistochemistry in uterine and ovarian carcinosarcomas and compared these findings to normal endometrium and ovarian control tissues. We have evaluated Trop-2 expression in multiple biologically aggressive, chemotherapy-resistant carcinosarcoma cell lines. Additionally, we have tested the sensitivity of these primary cell lines to immune-mediated cell death in the presence of hRS7, a humanized Trop-2 mAb made by grafting

the complementary-determining regions of its murine counterpart (mRS7) onto human IgG1 framework regions [11, 13–15]. To our knowledge, this is the first time that Trop-2 protein has been demonstrated to be significantly upregulated in human carcinosarcomas

from the uterus (UMMT) and ovary (OMMT), with negligible expression being detected in normal ovarian and uterine tissues. Significantly, Trop-2 positivity was confined to the epithelial component of the carcinosarcomas, without exception. Ixazomib Although the relationship between high Trop-2 expression and the aggressiveness of human epithelial neoplasms remains unclear, there is evidence that Trop-2 functions in the transduction of cell signals regulating tumor cell growth and resistance to apoptosis. Trop-2 possesses cytoplasmic serine and tyrosine phosphorylation sites and might function as a cell signal transducer and regulator of tumor cell growth while increasing tumor cell resistance to apoptosis [16]. Consistent with this, Trop-2 has been identified as an oncogene, implicated in colon cancer tumor growth, migration, and invasion, which suggests that Trop-2- specific targeting may inhibit tumor cell growth, migration and invasion [17]. Several human cancers have been shown to express a bicistronic CYCLIN D1-TROP2 mRNA chimera that acts as an oncogene and is able to induce aggressive tumor growth [18]. These observations support the possibility that aberrant Trop-2 expression contributes to the enhanced biologic aggressiveness of multiple human cancers, including carcinosarcomas.

Data extraction Two investigators independently reviewed the arti

Data extraction Two investigators independently reviewed the articles to exclude irrelevant and overlapping studies. The results were compared, and disagreements were resolved by discussion and consensus. When overlapping articles were found, we only included the publication that reported the most extensive information. From each study, the following information was extracted: journal, year of publication, first author, demographics, racial background CYT387 cost of the

study population, validity of the genotyping Selleck Saracatinib method, matching, and the number of cases and controls for each genotype. Frequencies of alleles were calculated for the cases and the controls, from the corresponding genotype distributions. Statistical analysis Review Manager 5.0 software

(The Cochrane Collaboration, Oxford, UK) was used for meta-analysis. The following genotype contrasts for the HIF-1α 1772 C/T polymorphism were evaluated: homozygotes TT versus a combination of CT and CC [TT versus (CT+CC), recessive model], a combination of TT and CT versus CC [(TT+CT) versus CC, dominant model]. Contrast of C allelic frequency versus G allelic frequency selleckchem (C versus G) was also evaluated. A allele of the HIF-1α 1790 G/A polymorphism was very rare. In most of the studies, homozygote AA was totally absent in both case and controls. For the HIF-1α 1790 G/A polymorphism, we only performed allelic frequency comparison (A versus G) and dominant model comparison [(AA+AG) versus GG]. In addition, we conducted subgroup analyses by cancer types, ethnicity, and gender. Etofibrate For gender subgroups, we included prostate cancer in male subgroup, and female specific cancers such as breast cancer, endometrial cancer, ovarian cancer and cervix cancer in female subgroup. We only conducted the meta-analysis on the subgroup with more than

two studies in Hardy-Weinberg equilibrium (HWE). For the HIF-1α 1790 G/A polymorphism, the pooled effects for other cancers (exclusion of the study on breast cancer) were also performed. The existence of heterogeneity between studies was ascertained by Q-statistic. The pooled odds ratio (OR) was estimated with models based on fixed effects or random effects assumptions. If the significant Q statistic (P < 0.1) indicated heterogeneity across studies, a random effects model was used for meta-analysis. Otherwise, a fixed effect model was selected. The 95% confidence interval (CI) of OR was also calculated. The distributions of genotypes in the controls were checked for HWE. Studies with the controls not in HWE were subjected to a sensitivity analysis [23]. The publication bias among the studies from the cases versus controls was assayed. Funnel plots of the HIF-1α 1772 C/T polymorphism for T versus C and HIF-1α 1790 G/A polymorphism for A versus G were performed to look for evidence of publication bias.

Figure 5 Analysis of anthramycin production by HPLC/MS After sep

Figure 5 Analysis of anthramycin production by HPLC/MS. After separating anthramycin on an HPLC column, mass spectrometry was performed using 6520 Agilent Accurate-Mass Q-TOF LC/MS. Conclusions This study shows that by isolation of new strains and testing

several plasmids, a host-vector system in a fast-growing and moderately thermophilic Streptomyces species could be developed. Two antibiotic biosynthetic gene clusters from mesophilic and thermophilic Streptomyces were heterlogously expressed in one strain. We expect that by utilizing thermophilic Streptomyces-specific promoters, more genes and especially antibiotic genes clusters of mesophilic Streptomyces should be heterologously expressed. Methods Bacterial strains, plasmids, JAK inhibitor and general methods Strains used in this work are listed in Table 1. Plasmid isolation, transformation of E. coli DH5α and PCR amplification followed Sambrook et al. [42]. 4SC-202 clinical trial Streptomyces culture, plasmid isolation and preparation of protoplasts and transformation of Streptomyces lividans ZX7 followed Kieser et al. [6]. Plasmid trans-conjugation from E. coli ET12567/pUZ8002 into thermophilic Streptomyces strains followed Bierman et al.

[38]. KpnI-treated pTSC1 was cloned in pBluescript II SK to obtain pCWH100 and was sequenced by primer-walking at Shanghai Invitrogen Inc. Sequence comparisons were done with software from the National Center for Biotechnology Information http://​www.​ncbi.​nlm.​nih.​gov/​BLAST. The complete nucleotide sequence of pTSC1 was deposited in the GenBank database under no. GU271942. Isolation and identification of thermophilic Streptomyces strains Samples of P505-15 cost garden soil, weed compost and swine manure were collected from Shanghai city, Hunan, Hubei and Fujian provinces in the summers of 2005 and 2006. The samples 4-Aminobutyrate aminotransferase were dried at 100°C for 1 h and cultivated on SC medium (starch 10 g, casein 0.3 g, KNO3 2 g, MgSO4.7H2O 0.05 g, FeSO4.7H2O 0.01 g, CaCO3 0.02 g, agar 18 g, H2O to 1000 ml, pH7.2) [43] at 50°C for 3-5 d. Thermophilic Streptomyces strains were cultured in TSB (Oxoid tryptone soya broth powder, 30 g, H2O to 1000 ml)

liquid medium at 45°C for 1 d and genomic DNA was isolated followed the Kirby mix procedure [6]. 16S rRNA genes were amplified by PCR with primers (5′-AGAGTTTGATCCTGGCTCAG-3′ and 5′-TCAGGCTACCTTGTTACGACTT-3′). PCR conditions were: template DNA denatured at 95°C for 5 min, then 95°C 30 s, 55°C 30 s, 72°C 2 min, for 35 cycles. PCR products were cloned in pBluescript II SK and sequenced with its T7 and T3 primers. Strains were inoculated on MS (mannitol 20 g, soya flour 20 g, agar 20 g, H2O to 1000 ml, pH7) medium covered with cellophane disks. After 2 days incubation at 42°C, the cells were fixed with fresh 2% glutaraldehyde (pH7.2) and 1% osmium tetroxide. Spores were examined with a JSM-6360LV scanning electron microscopy (Jeol).

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