All cleaned substrates were treated with UV Ozone treatment for 15 min. Figure 1 Detailed values extracted from the UPS spectra and schematic diagram of organic solar cells. (a) Evolution of secondary electron edge of ITO and ITO/ZnOCs2CO3 and (b) energy level alignment of all materials used in this study. The solution for electron selective layer was prepared by mixing ZnO and Cs2CO3 with different blend ratios, namely, 1:1, 1:2, 1:3, 2:1, and 3:1. The solution-processed ZnO or ZnO:Cs2CO3 was spin-coated at 1,000 rpm for 25 s onto the cleaned substrates and later annealed at 300°C for 10 min. The photoactive layer either P3HT:PCBM or P3HT:ICBA dissolved in 1,2-dichlorobenzene

was spin-coated at 700 rpm for 25 s and subsequently annealed at 130°C for 30 min or 150°C for 10 min, respectively. FK866 datasheet Later, PEDOT:PSS was spin-coated at 4,000 rpm for 25 s onto the photoactive layer and annealed at 120°C for 20 min. To complete the device, 100-nm thick of Al was thermally selleck inhibitor selleck screening library evaporated at rates 4 A/s through a shadow mask at a base pressure of 10−7 Torr. The active area of the complete devices is 0.04 cm2. To ensure the reproducibility of our results,

we have fabricated 83 devices throughout this work. The following are the fabricated devices based on different photoactive materials. P3HT:PCBM-based devices. Device A-ITO/ZnO/P3HT:PCBM/PEDOT:PSS/Al Device B-ITO/ZnO:Cs2CO3/P3HT:PCBM/PEDOT:2PSS/Al P3HT:ICBA-based devices. Device C-ITO/ZnO/P3HT:ICBA/PEDOT:PSS/Al Device D-ITO/ZnO:Cs2CO3/P3HT:ICBA/PEDOT:PSS/Al Thin film and device characterizations The J-V characteristics of the conventional solar cells were measured using the Keithley 2400 source meter under a solar simulator (AM1.5) with an irradiation intensity of 100 mW/cm2. The EQE measurements were performed using an EQE system (Model 74000) obtained from Newport Oriel Instruments, Irvine, CA, USA, and the HAMAMATSU calibrated silicon cell photodiode (HAMAMATSU, Shizuoka, Japan) was used as the 4��8C reference diode. The wavelength was controlled with a monochromator to range from 200 to 1,600 nm. AFM imaging

was achieved in air using a Digital Instrument Multimode that is equipped with a nanoscope IIIa controller. XPS measurements were performed in a PHI 5000 VersaProbe (Ulvac-PHI, Chigasaki, Kanagawa, Japan) with background pressure of 6.7 × 10−8 Pa, using a monochromatized Al Kα (hv = 1,486.6 eV) anode (25 W, 15 kV). Ultraviolet photoemission spectroscopy (UPS) measurements were carried out using the He 1 photon line (hv = 21.22 eV) of a He discharge lamp under UHV conditions (4 × 10−10 mbar). The transmittances of ZnO, and ZnO:Cs2CO3 coated on ITO-glass substrates were recorded at room temperature with a SCINCO S4100 (SCINCO, Seoul, South Korea) spectrophotometer. XRD measurements were carried out using X’PERT PRO of PANalytical Diffractometer (PANalytical, Seongnam City, South Korea) with a Cu Kα source (wavelength of 1.

Sequencing was performed using ACP-196 cost ABI310 Genetic Analyser (Applied Biosystems), and data were collected using ABI Prism 310 Data Collection Software. Results and discussion All the positive and negative controls used in this study were selected by Sanger sequencing of patients’ samples. The results obtained using endonuclease

restriction, ARMS and HRM were verified with those obtained using Sanger sequencing to determine the specificity of the assays. Sensitivity was measured as the minimal percentage of mutated allele in a sample detected by the assay. The initial portion of mutation was determined using Sanger sequencing. DNMT3A mutation analysis Endonuclease restriction analysis identified DNMT3A R882H G>A find more mutations in 28 out of 230 patients with AML (12.2%) and HRM analysis identified 2 additional R882X G>C mutations (0.9%), which are consistent with the frequency published by Lin et al. [28]. The age of the patients ranged from 24 to 87 years (median, 58 years). Among these patients, 53% had a normal karyotype. None of the patients in the prognostic favourable group had DNMT3A mutations. Of 30 patients, 16 had FLT3 mutations. Figure 1 provides a representative result of restriction analysis with 5 positive this website and 2 negative samples. Point mutation at R882H (GCCGC to GCCAC) led to the loss of

one recognition site of Fnu4HI, thus creating a larger 289 bp

fragment. Because of heterozygosity, the 190 bp wt fragment and the smaller 114 bp fragment are present in every sample. Sensitivity of the assay was analysed using serial dilutions of wt and DNMT3A R882H-mutated DNA (initial mutation ratio in Sanger sequencing was 59%, Figure 2.1). The fragment containing the mutation was explicitly apparent with a mutational content of 0.05%, indicating a very high sensitivity of the assay. In addition mutations in exon 23 of DNMT3A were detected using HRM analysis. Results of HRM analysis were plotted as a difference in the fluorescence of the tested sample versus that of a wt control (normalisation line), referred to as a temperature-shifted difference plot (Figure 3.1). Discrepancies between mutated and wt samples could also be observed in the melting plot profiles. Sample containing R882H mutation ADAMTS5 showed 2 peaks at 84.5°C and 85.6°C, whereas the wt samples showed only 1 peak at 85.7°C. Compared to the wt allele, R882X allele was slightly shifted to the left, with a melting temperature of 85.6°C (Figure 3.2). Sensitivity of the HRM assay was assessed similar to that of restriction analysis. The assay had high confidence (97%-99%) for the mutated allele up to a mutation ratio of 5.9% (Figure 2.2). Lower mutation ratios could not be assigned as positive and were identified as false negative with a confidence of 92%-98%.

Design of the

studies differed with variation in recruitment methods and inclusion criteria. All patients had to have had a biopsy (from inclusion criteria) which could introduce verification bias compared to those patients with excess alcohol consumption not selected for biopsy having a different disease severity than those who were selected. Only four studies reported any parameters by which biopsy quality could be judged, and half of these reported findings stratified by biopsy quality. Even when the tests were similar between studies, the thresholds used were different or not reported. Direct comparison between studies was made more difficult by the use of a range of fibrosis staging systems, largely locally generated. There was heterogeneity learn more and lack of standardization of analytical methods used for the markers measurements and as these different assays may not be well correlated, external validity may be reduced and the determination of a single generalisable threshold remains problematic for those markers assayed locally. Access and availability of serum markers using commercial automated platforms may address this issue. There was incomplete reporting of co-morbidities and diagnostic

test results, making appraisal and summative assessment difficult. The paucity of studies which looked at direct comparisons between panels, BIBF 1120 clinical trial and between single marker and panels make it difficult to C-X-C chemokine receptor type 7 (CXCR-7) say one panel is more accurate than another. It is clear from this systematic Selleckchem MLN8237 review that the current serum markers are promising, improving and may provide additional diagnostic information in the identification and management of people with ALD. The limitations of this review include lack of data to perform summative analyses and a focus on the ability of diagnostic tests to identify fibrosis alone. Detection of inflammation has not been addressed. Issues of spectrum bias which may have an impact on performance

characteristics of the tests making direct comparisons between studies problematic, and this has not been directly addressed in this review. This is due to several main problems in accounting for such as bias. The first is a lack of a universally accepted system of dealing with this issue, especially in this group of patients with ALD. There have been some methodological suggestions published by one group in chronic Hepatitis C [39], who have used this method in a study in ALD patients [30]. Authors used standard population of same prevalence for all fibrosis stages and currently it is unclear if this has external validity or international acceptance by professionals working in this field. In addition the studies included in this review are older, use different classification systems for histology and have inconsistent and incomplete reporting of the individual stages of study participants.

melanogaster w1118 . This increase is possibly caused by the specific effect of the Wolbachia strain wMelPop, since it was not observed in wMel-infected D. melanogaster Canton S. Our current electron microscopic observations allowed us to identify

changes in Wolbachia morphology in apoptotic germline cells. Morphological evidence of apoptosis in germarium cells The ultrastructural features of apoptosis in the cyst cells of higher eukaryotes have gained wide recognition. They include cytoplasmic and nuclear condensation (pyknosis); nuclear fragmentation (karyorrhexis); normal morphological appearance of cytoplasmic organelles; see more an intact plasma membrane [3, 4]. The ultrastructural AICAR changes we identified here in D. melanogaster cyst cells are consistent with the above hallmarks. Furthermore, we revealed mitochondria of two types: intact morphology in one type and markedly swollen with a few cristae in the other. A similar heterogeneity of mitochondrial ultrastructure has been observed during apoptosis in granulose cells of Japanese quail (Coturnix coturnix japonica) [30], lymphocytes from leukemia patients [31],

and megakaryocytes from patients with idiopathic thrombocytopenic purpura [32]. It has been suggested that the swollen mitochondria release cytochrome c, which activates a cascade of proteolytic reactions, while the normal ones retain their capacity for ATP synthesis, a process apoptosis requires [30, 31, 33]. According to our qualitative analysis using EM, morphological evidence of apoptosis was revealed in germline cells from uninfected flies and those infected with wMel and wMelPop. Thus, there are this website reasons for inferring that the endosymbiont Wolbachia in D. melanogaster cystocytes has no effect on sequential passage of intracellular organelles through apoptosis. To reveal the possible differences

between the effect of the wMel and wMelPop strains on apoptosis in the germaria, additional Megestrol Acetate morphometric analysis of the number of apoptotic structures and of Wolbachia density in the cystocytes is required. Structural features of Wolbachia in apoptotic cysts Wolbachia with matrix of moderate and low electron density in apoptotic cells in region 2a/2b of the germarium have been previously encountered in other types of D. melanogaster ovaries [34] and they presumably reflect different functional states of bacteria. Wolbachia with disrupted envelopes and light matrix are possibly dying bacteria in apoptotic cells. Such appearance has not been observed in Wolbachia injured or killed by heat stress [35] and tetracycline [36]. The electron-dense bacteria-like structures at the periphery of region 1 of the germarium may be evidence of changes in dying Wolbachia.

Downstream from this region, a very high divergence was observed with 37,6%, 37,8%, and 38,7% aa identity, respectively. Likewise, in this region, MS2/28.1 shared only 39,8% and 38,8% identity, respectively, with the two vlhA1 expressed variants, vlhA4 and vlhA5, previously identified in M. synoviae strain WVU 1853 (Figure 2). Overall, the haemagglutinin region of MS2/28.1

was found to be considerably reduced in size (148 aa less than in vlhA1) and displayed high level of sequence divergence in comparison to the previously reported vlhA expressed genes, namely vlhA1, vlhA4 (GenBank accession no. AF181033), Selleckchem BIIB057 and vlhA5 (GenBank accession no. AF181034) [17]. Figure 2 Comparison of the amino acid sequence predicted from M. synoviae MS2/28.1 gene with vlhAs 1 to 5. Alignment of the completed full-length MS2/28.1 deduced amino acid sequence with vlhAs 1 to 5 (GenBank accession numbers AF035624, AF085697, AF085698, AF181033, and AF181034, respectively). Identical aa regions are selleck kinase inhibitor shaded in black while similar aa residues are shaded in grey. Demonstration that MS2/28.1 sequence is preceded by the vlhA1 promoter To confirm that in our bacterial

stock MS2/28.1 was located downstream of the unique vlhA promoter sequence, we performed PCR amplifications on single colonies using oligonucleotide primers placed in the vlhA promoter sequence with either vlhA1- or MS2/28.1-specific reverse primers. As shown in Figure 3, amplicons migrating at the expected mobility were obtained solely with MS2/28.1-specific reverse primers. Sequence analysis selleck chemical further confirmed that the upstream sequence is identical to that of the vlhA1 promoter, a result consistent with the finding that MS2/28.1 is transcriptionally active and that, in its transcript, the region preceding its ATG initiation codon was identical to that reported for vlhA1. Figure 3 Confirmation

that MS2/28.1 is preceded by the unique vlhA1 promoter sequence. Primer EXpro, which anneals to the vlhA1 promoter, was combined with either vlhA1R (lanes b) or with ORF5.1R (lanes c). No amplification from genomic DNA extracted from the four colonies was obtained with the vlhA1-specific reverse primer (lanes b). Expected amplicon was obtained with primers EXpro/ORF5.1R (lanes c). PCR amplification of the full length MS2/28.1 was Vorinostat in vivo obtained with the primers pair EXproint and 2/28.1Rev (lanes d). As negative control, PCR was performed with no genomic M. synoviae DNA (lane a). Lane M; DNA size marker (1 kb). MS2/28.1 encoded full-length product is post-translationally cleaved with its C-terminal portion exposed at the bacterium’s surface To characterize MS2/28.1 encoded product and to examine whether it was processed similarly as the vlhA1 product, we generated antisera towards four bacterially expressed distinct regions of the coding sequence. The reactivity of these antisera is shown in Figure 4.

In addition, the indicator phenol red was added to all wells of the Taxa Profile™ A and C microtiter plates to optimize detection. The blank value was measured for each biochemical reaction on the same plate and subtracted from measured values. In order to assess inter-assay variability five independent experiments per strain were conducted. For evaluation of the newly developed Brucella specific 96-well microtiter plate three trials

per strain were run independently. Intra-assay variability was assessed with the reference strains testing all substances twice within the same experiment. Since the blank values measured on extra plates proved to be constant a fixed mean value of each substrate was subtracted from the measured data. Data acquisition and analysis Turbidity and colour change were measured photometrically using a Multiskan Ascent® photometer LY2606368 in vivo (Labsystems,

Helsinki, Finland) at a wave length of 405 nm, 540 nm and 620 nm according to manufacturer’s recommendations. Optimal OD cut-off values were empirically adapted from the preliminary test results of the 384-wells Taxa Profile™ microtiter plates. Stable and discriminatory markers were selected to design a 96-well Micronaut™ plate (Figure 2) to identify bacteria of the genus Brucella and to classify their species and biovar. Dendrograms were deduced from Erastin nmr the biotyping data using SPSS version 12.0.2 (SPSS Inc., Chicago, IL, USA). First of all, three different character data sets were defined following

the metabolic activity tested (Taxa Profile™ A (“”amino acids”"), C (“”carbohydrates”"), and E (“”other enzymatic reactions”")). Each character was considered as equal within the particular data set. Both the raw OD data and the binary coded data based on the empirically set cut-off were analyzed using the Pearson coefficient and the categorical coefficient, respectively. Hierarchical cluster analysis was performed by the Ward’s linkage algorithm, and a dendrogram was generated. If necessary, analysis was repeated within each cluster for further discrimination. Secondly, a separate data analysis Interleukin-3 receptor of the 23 Brucella reference strains representing the currently known species and biovars was performed including all biochemical reactions of the Taxa Profile™ system or exclusively the TPCA-1 clinical trial substrates selected for the newly developed plate. Finally, the whole collective of 113 strains tested with the Brucella specific Micronaut™ microtiter plate was analyzed to prove the diagnostic system. An identification table presenting quantitative and qualitative metabolic activity was created [Additional file 7] and the specificity of the test system to differentiate Brucella species and biovars was calculated (Table 1). Acknowledgements The project was partially supported by research funds of the Bundeswehr Medical Service. We are grateful to Dr.

S. National Herbarium 36(3):63–119 Wirth M, Hale ME Jr (1978) Morden-Smithsonian Expedition to Dominica: the lichens (Graphidaceae). Smithson Contrib Bot 40:1–64CrossRef”
“Introduction Species of genus Myceliophthora and its teleomorph Corynascus have attracted increasing interest due to their potential to produce thermostable enzymes. For instance, laccases of M. thermophila (basionym: Sporotrichum thermophilum) were shown to be thermostable

with high activity after expression in different expression hosts (Berka et al. 1997; Bulter et al. 2003; Babot et al. 2011). Due to the potential of Myceliophthora to degrade lignocellulolytic plant material, many (hemi-)cellulolytic enzymes of M. thermophila are characterized and patented

(Bhat and Maheshwari 1987; Roy et al. 1990; Sadhukhan et al. 1992; Badhan et al. 2007; Beeson et al. 2011). The importance of this fungal Lazertinib chemical structure group has recently been underlined by the sequencing of the genome of M. thermophila isolate ATCC42464 (genome.jgi-psf.org/Spoth1). The first Myceliophthora species, M. lutea, was described by Constantin and Matruchot in 1894 as a Cytoskeletal Signaling inhibitor pathogen causing the ‘vert de gris’ mat disease of cultured mushrooms (Costantin 1892). This species was classified before as a member of the genus Chrysosporium (Carmichael 1962), but there after von Arx re-introduced the genus Myceliophthora and its type species M. lutea (von Arx 1973). Initially, three species were assigned to this genus: M. fergusii, M. lutea, and M. thermophila (van Oorschot 1980). Another species, M. vellerea, was most likely wrongly described as a Myceliophthora species based on morphological differences selleck products (Sigler et al. 1998). A fourth species, M. hinnulea, was assigned to the genus Myceliophthora by Awao and Udagawa (1983). The type species of the ascomycete genus Corynascus, C. sepedonium, was described by Emmons (1932). This species was originally part of the genus Thielavia before von Arx introduced the genus Corynascus. This genus can be distinguished from Thielavia by the presence of SDHB ascospores with two germ pores, one at each end (von Arx 1973). At that time, the genus Corynascus contained the species C. sepedonium and C. novoguineensis (von Arx

1973 ). Currently, seven Corynascus species are described: C. heterothallicus, C. novoguineensis, C. sepedonium, C. sexualis, C. similis, C. thermophilus and C. verrucosus (von Klopotek 1974; Stchigel et al. 2000). Of all the species of these two genera, M. thermophila is most commonly used for applied research (Roy et al. 1990; Berka et al. 1997; Rosgaard et al. 2006; Badhan et al. 2007; Beeson et al. 2011). Several isolates of M. thermophila can grow at temperatures up to 50°C on cellulose-rich material and can decompose complex substrates such as birch chips, wood pulp and wheat straw (Bhat and Maheshwari 1987). M. thermophila was initially classified in the genus Sporotrichum (Fergus and Sinden 1969) before it was assigned to the genus Chrysosporium as C.

Thin Solid Films 1993, INCB018424 manufacturer 236:27–31.CrossRef 17. Lou XC, Zhao XJ, He X: Boron doping effects in electrochromic properties of NiO films prepared by sol–gel. Sol Energy 2009, 83:2103–2108.CrossRef 18. Steinebach H, Kannan S, Rieth L, Solzbacher F: H 2 gas sensor performance of NiO at high temperatures in gas mixtures. Sensors Actuators B-Chem 2010, 151:162–168.CrossRef

19. Adler D, Feinleib J: Electrical and optical properties of narrow-band materials. Phys Rev B-Solid State 1970, 2:3112–3134.CrossRef 20. Chung JL, Chen JC, Tseng CJ: Preparation of TiO 2 -doped ZnO films by radio frequency magnetron sputtering in ambient hydrogen–argon gas. Appl Surf Sci 2008, 255:2494–2499.CrossRef 21. Kang JK, Rhee SW: Chemical vapor deposition of nickel oxide films from Ni(C 5 H 5 ) 2 /O 2 . Thin Solid Films 2001, 391:57–61.CrossRef 22. Zheng K, Gu L, Sun D, Mo XL, Chen G: The properties of ethanol

gas sensor based on Ti doped ZnO nanotetrapods. Mater Sci Eng B 2009, 166:104–107.CrossRef 23. Reguig BA, Khelil A, Cattin L, Morsli M, Bernède JC: Properties of NiO thin films deposited CHIR98014 by intermittent spray pyrolysis process. Appl Surf Sci 2007, 253:4330–4334.CrossRef 24. Pala RGS, Tang W, Sushchikh MM, Park JN, Forman AJ, Wu G, Kleiman-Shwarsctein A, Zhang J, McFarland EW, Metiu H: CO oxidation by Ti- and Al-doped ZnO: oxygen activation by adsorption on the SCH727965 molecular weight dopant. J Catal 2009, 266:50–58.CrossRef 25. Burstein E: Anomalous optical absorption limit in InSb. Phys Rev 1954, 93:632–633.CrossRef 26. Hamberg I, Granqvist CG,

Berggren KF, Sernelius BE, Engstrom L: Band-gap widening in heavily Sn-doped In 2 O 3 . Phys Rev B 1984, 30:3240–3249.CrossRef 27. Serpone N, Lawless D, Khairutdinov R: Size effects on the photophysical PLEKHB2 properties of colloidal anatase TiO 2 particles: size quantization versus direct transitions in this indirect semiconductor. J Phys Chem 1995, 99:16646–16654.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions C-C H carried out the experimental procedures, including the depositions of NiO and TZO thin films and measurements of SEM and X-ray patterns. F-H W gave the suggestion for the paper organization and English grammar correction. C-F Y participated in the design of the study, performed the statistical analysis, and organized the paper. C-C W and H-H H participated in the measurement and prediction of the I-V curve of NiO/TZO heterojunction diodes using the space-charge limited current (SCLC) theorem. All authors read and approved the final manuscript.”
“Background Silicon oxynitride (SiO x N y ) is a very useful material for applications in microelectronic and optoelectronic devices due to the possibility of tailoring the film composition and property according to the O/N ratio.

Universal PCR primer designed for NTNH Sequences used to design a set of universal

PCR primers were MM-102 mouse obtained from Genbank. All sequences were aligned with MegAlign (DNASTAR, Lasergene, Inc.). Both Primer Express (Applied Biosystems, Foster City, CA) and Primer3 (http://​frodo.​wi.​mit.​edu/​primer3/​), were used to design a pair of degenerate primers that included base differences to detect all known NTNH gene variants. Primer sequences are designated in Figure 1. Universal PCR for detection of NTNH of all C. botulinum types Purified DNA from C. botulinum, E. coli bacterial DNA (pUC19 plasmid DNA) or crude lysate from human leukocytes were used in the universal

PCR. PCR conditions were as follows: 95°C for 5 minutes, then 35 cycles of 95°C for 15 seconds and 57°C for 1 minute. PCR reaction mixture contained PCR Buffer, Adavosertib mw 3.5 uM MgCl2, 200 nM dNTP, 1 uM forward or reverse primer, 0.25 U Taq Polymerase (Invitrogen Corp, Carlsbad, CA). 5 μL of DNA (0.25 ng/uL) was used in each 25 μL PCR reaction. PCR products selleck screening library were run on a 2.5% agarose gel to separate the product from any non-specific amplification and visualized for 101 bp bands by UV illumination. Toxin type-specific qPCR primer and probe design Neurotoxin gene sequences, obtained both from Genbank and from sequences provided by Biosciences Division, Los Alamos National Laboratories, were aligned and degenerate primer/probe sets were designed using software packages as above for each toxin type. Each degenerate primer/probe set include all known base differences within each toxin type. Generation of qPCR standards for each C. botulinum toxin type-specific assay Seven samples of purified C. botulinum DNA, one for each toxin type, were

used in the generation of plasmid DNA standards for qPCR. Briefly, primers designed specifically for each toxin type were used to amplify a region of the toxin gene containing Non-specific serine/threonine protein kinase the degenerate primer/probe set target sequences. The PCR conditions were as follows: 95°C for 5 minutes, then 35 cycles of 95°C for 15 seconds and 60°C for 1 minute. PCR reaction mixture contained PCR Buffer, 3.5 uM MgCl2, 200 nM dNTPs, 500 nM forward or reverse primer, 0.25 U Taq Polymerase (Invitrogen). 5 μL of DNA (0.25 ng/uL) was used in each 25 μL PCR reaction. PCR products were visualized by UV on a 1.5% agarose gel. Corresponding specific products were gel purified and ligated into pGEM T-easy vector (Promega Corp., Madison, WI). Ligations were transformed into DH5α E.coli bacteria using α-complementation to determine positive colonies. Positive colonies were grown in overnight cultures, plasmid DNA was purified and sequenced for determination of correct subtype insert sequence.

Methods Bacterial strains and growth conditions The strains and selleck plasmids used in this study are described in Additional

file 1: Table S2. C. crescentus strains were cultured at 30°C in M2 minimal salts medium plus glucose [39]. When appropriate, the growth medium was supplemented with chloramphenicol (1 μg ml-1), kanamycin (10 μg ml-1) or tetracycline (2 μg ml-1). Plasmids were propagated in Escherichia coli strain DH5α (Invitrogen) and mobilized into C. crescentus by bacterial conjugation using E. coli strain S17-1 [40]. E. coli strains were grown at 37°C in LB broth [41]. Deletion of genes CC2906 Androgen Receptor inhibitor and CC3255 in C. crescentus Single mutant strains for CC2906 (SG20) and CC3255 (SG19) were obtained by an in-frame deletion in the coding region of these genes. For that, two fragments flanking the regions to be deleted were amplified by PCR (a complete list of primers used in this study is in Additional file 1: Table S3) and subcloned into pNPTS138 [42]. Constructs into pNPTS138 were transferred to C. crescentus strain NA1000

[43] by conjugation with E. coli S17-1 and the deletion of the wild-type copy of the gene in the NA1000 background was achieved by two homologous recombination events. Mutant strains were isolated by screening colonies by PCR and DNA sequencing. For the construction of a double mutant strain

(SG21), the single mutant strain SG20 was used for the two homologous recombination events of the CC3255 deletion. Construction of point mutations in CC3252 and overexpression of CC3252 in C. Arachidonate 15-lipoxygenase crescentus Codons for the conserved cysteine residues of the protein encoded by CC3252 (C131 and C181) were replaced for a codon corresponding to Blasticidin S serine by overlapping PCR with a pair of complementary primers (Additional file 1: Table S3) designed for each substitution. Each part of CC3252 was amplified separately by PCR using one of each complementary primer set and a primer hybridizing upstream or downstream from CC3252. The partially complementary PCR products were used together as templates in a second amplification reaction with the primers hybridizing upstream and downstream from CC3252. The amplicons obtained were cloned into pGEM-T (Promega) and sequenced. The inserts were excised from vectors and subcloned into pNPTS138. Constructs into pNPTS138 were transferred to C. crescentus strain NA1000 [43] by conjugation with E. coli S17-1 and replacement of the wild-type copy of the gene for the corresponding mutated copy in the NA1000 background was achieved by two homologous recombination events.