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https://www.readbyqxmd.com/read/28335553/biosynthesis-of-polyunsaturated-fatty-acids-in-octopus-vulgaris-molecular-cloning-and-functional-characterisation-of-a-stearoyl-coa-desaturase-and-an-elongation-of-very-long-chain-fatty-acid-4-protein
#1
Óscar Monroig, Rosa de Llanos, Inmaculada Varó, Francisco Hontoria, Douglas R Tocher, Sergi Puig, Juan C Navarro
Polyunsaturated fatty acids (PUFAs) have been acknowledged as essential nutrients for cephalopods but the specific PUFAs that satisfy the physiological requirements are unknown. To expand our previous investigations on characterisation of desaturases and elongases involved in the biosynthesis of PUFAs and hence determine the dietary PUFA requirements in cephalopods, this study aimed to investigate the roles that a stearoyl-CoA desaturase (Scd) and an elongation of very long-chain fatty acid 4 (Elovl4) protein play in the biosynthesis of essential fatty acids (FAs)...
March 21, 2017: Marine Drugs
https://www.readbyqxmd.com/read/28335389/characteristics%C3%A2-and%C3%A2-lethality%C3%A2-of%C3%A2-a%C3%A2-novel%C3%A2-recombinant%C3%A2-dermonecrotic%C3%A2-venom%C3%A2-phospholipase%C3%A2-d%C3%A2-from%C3%A2-hemiscorpius%C3%A2-lepturus
#2
Elham Torabi, Mahdi Behdani, Mohammad Hosseininejad Chafi, Reza Moazzami, Jean-Marc Sabatier, Vahid Khalaj, Delavar Shahbazzadeh, Kamran Pooshang Bagheri
Hemoscorpius lepturus is the most medically important scorpion in Iran. The clinical signs of H. lepturus envenomation are remarkably similar to those reported for brown spiders, including dermonecrosis, hematuria, renal failure and even death. The lethality and toxicity of brown spiders' venom have been attributed to its phospholipase D activity. This study aims to identify a phospholipase D with possible lethality and dermonecrotic activity in H. lepturus venom. In this study, a cDNA library of the venom glands was generated by Illumina RNA sequencing...
March 13, 2017: Toxins
https://www.readbyqxmd.com/read/28334760/twin-primer-non-enzymatic-dna-assembly-an-efficient-and-accurate-multi-part-dna-assembly-method
#3
Jing Liang, Zihe Liu, Xi Z Low, Ee L Ang, Huimin Zhao
DNA assembly forms the cornerstone of modern synthetic biology. Despite the numerous available methods, scarless multi-fragment assembly of large plasmids remains challenging. Furthermore, the upcoming wave in molecular biological automation demands a rethinking of how we perform DNA assembly. To streamline automation workflow and minimize operator intervention, a non-enzymatic assembly method is highly desirable. Here, we report the optimization and operationalization of a process called Twin-Primer Assembly (TPA), which is a method to assemble polymerase chain reaction-amplified fragments into a plasmid without the use of enzymes...
February 23, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28334704/combining-real-time-cold-and-mama-pcr-taqman-techniques-to-detect-and-quantify-r201-gnas-mutations-in-the-mccune-albright-syndrome%C3%A2
#4
Luisa de Sanctis, Ilaria Galliano, Paola Montanari, Patrizia Matarazzo, Daniele Tessaris, Massimiliano Bergallo
BACKGROUND/AIM: The McCune-Albright syndrome (MAS) is a potentially severe disorder hallmarked by fibrous bone dysplasia, café-au-lait skin spots, and endocrine hyperfunction. It is caused by postzygotic activating mutations at the R201 codon of the GNAS gene, leading to a state of somatic mosaicism. Our aim was to improve the mutation detection rate and to quantify the presence of R201 GNAS mutations in different DNA samples from MAS patients. METHODS: Real-time COLD- and MAMA-PCR TaqMan techniques were combined to search for R201 mutations in the DNA of blood or affected tissues from 16 previously molecularly characterized MAS patients, from a further 84 subjects with MAS signs who were R201 negative at RFLP analysis, and from 36 controls...
March 23, 2017: Hormone Research in Pædiatrics
https://www.readbyqxmd.com/read/28334619/investigation-on-metal-tolerance-and-phytoremoval-activity-in-the-poplar-hybrid-clone-monviso-under-cu-spiked-water-potential-use-for-wastewater-treatment
#5
Fabrizio Pietrini, Daniela Di Baccio, Valentina Iori, Sofia Veliksar, Natalia Lemanova, Liudvika Juškaitė, Audrius Maruška, Massimo Zacchini
A serious concern for the environmental and human health is represented by the increasing copper (Cu) occurrence in agricultural soils and waters, because of the possible food contamination and bioaugmentation along the trophic chain. The request for the decontamination of different matrices with an environmentally sustainable technology as the phytoremediation should be addressed by selecting plant materials with improved pollutant tolerance and removal capability. With this purpose, plants of the hybrid poplar clone "Monviso" (Populus×generosa A...
March 20, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28334280/origin-and-genetic-differentiation-of-pink-flowered-sorbus-hybrids-in-the-western-carpathians
#6
Veronika Uhrinová, Judita Zozomová-Lihová, Dana Bernátová, Juraj Paule, Ladislav Paule, Dušan Gömöry
Background and Aims: Diversity of the genus Sorbus has been affected by interspecific hybridizations. Pink-flowered hybrid species have been insufficiently studied so far. They comprise bigenomic hybrid species derived from crosses S. aria s.l. × S. chamaemespilus and trigenomic ones, where S. aucuparia was involved as well. The main objective of the present study was to reconstruct their hybrid origins as well as to assess genetic distinction among several morphologically recognized hybrid species...
March 13, 2017: Annals of Botany
https://www.readbyqxmd.com/read/28334274/interactive-effects-of-multiple-climate-change-factors-on-ammonia-oxidizers-and-denitrifiers-in-a-temperate-steppe
#7
Cuijing Zhang, Jupei Shen, Yifei Sun, Juntao Wang, Limei Zhang, Zhongling Yang, Hongyan Han, Shiqiang Wan, Jizheng He
Global climate change could have profound effects on belowground microbial communities and subsequently affect soil biogeochemical processes. The interactive effects of multiple co-occurring climate change factors on microbially-mediated processes are not well understood. A four-factorial field experiment with elevated CO2, watering, nitrogen (N) addition and night warming was conducted in a temperate steppe of northern China. Real-time polymerase chain reaction and terminal-restriction fragment length polymorphism, combined with clone library techniques were applied to examine the effects of those climate change factors on N-related microbial abundance and community composition...
March 15, 2017: FEMS Microbiology Ecology
https://www.readbyqxmd.com/read/28334229/functional-and-nonfunctional-forms-of-cquior91-an-odorant-selectivity-subunit-of-culex-quinquefasciatus
#8
David T Hughes, Julien Pelletier, Suhaila Rahman, Sisi Chen, Walter S Leal, Charles W Luetje
In Culex quinquefasciatus, CquiOR91 is the ortholog of 2 larvae-specific odorant receptors (ORs) from Anopheles gambiae (Agam\Or40, previously shown to respond to several odorant ligands including the broad-spectrum repellent N,N-diethyl-3-methylbenzamide, DEET) and Aedes aegypti (Aaeg\Or40). When we cloned full-length CquiOR91 from a Culex quinquefasciatus larval head RNA sample, we found 2 alleles of this OR, differing at 9 residues. Functional analysis using the Xenopus oocyte expression system and 2-electrode voltage clamp electrophysiology revealed one allele (CquiOR91...
March 3, 2017: Chemical Senses
https://www.readbyqxmd.com/read/28333957/immune-response-of-rats-vaccinated-orally-with-various-plant-expressed-recombinant-cysteine-proteinase-constructs-when-challenged-with-fasciola-hepatica-metacercariae
#9
Malgorzata Kesik-Brodacka, Agnieszka Lipiec, Monika Kozak Ljunggren, Luiza Jedlina, Katarzyna Miedzinska, Magdalena Mikolajczak, Andrzej Plucienniczak, Andrzej B Legocki, Halina Wedrychowicz
BACKGROUND: Cysteine proteinases of Fasciola hepatica are important candidates for vaccine antigens because of their role in fluke biology and host-parasite relationships. In our previous experiments, we found that a recombinant cysteine proteinase cloned from adult F. hepatica (CPFhW) can protect rats against liver fluke infections when it is administered intramuscularly or intranasally in the form of cDNA. We also observed considerable protection upon challenge following mucosal vaccination with inclusion bodies containing recombinant CPFhW produced in Escherichia coli...
March 23, 2017: PLoS Neglected Tropical Diseases
https://www.readbyqxmd.com/read/28333450/pcr-independent-method-of-transformation-associated-recombination-reveals-the-cosmomycin-biosynthetic-gene-cluster-in-an-ocean-streptomycete
#10
Charles B Larson, Max Crüsemann, Bradley S Moore
The transformation-associated recombination cloning methodology facilitates the genomic capture and heterologous expression of natural product biosynthetic gene clusters (BGCs). We have streamlined this procedure by introduction of synthetic DNA gene blocks for the efficient capture of BGCs. We show the successful capture and expression of the aromatic polyketide antitumor agent cosmomycin from streptomycete bacteria and the discovery of new cosmomycin analogues by mass spectral molecular networking.
March 23, 2017: Journal of Natural Products
https://www.readbyqxmd.com/read/28333351/ecological-impact-of-ciprofloxacin-on-commensal-enterococci-in-healthy-volunteers
#11
Victoire de Lastours, Elena Maugy, Vincent Mathy, Françoise Chau, Benjamin Rossi, François Guérin, Vincent Cattoir, Bruno Fantin
Background: The ecological impact of ciprofloxacin on commensal enterococci is unknown. Methods: Forty-eight healthy volunteers received ciprofloxacin from day (D) 0 to D14; stools were collected on D7, D14 and D42. Fluoroquinolone-susceptible and -resistant enterococci (FQ-SE and FQ-RE) were detected and quantified by culture, and identified by MALDI-TOF MS. The relative abundance of FQ-RE over FQ-SE was determined. The genetic basis of fluoroquinolone resistance was deciphered by partial sequencing of gyrA and parC genes...
March 4, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/28333348/promoter-characterization-and-expression-of-the-blakpc-2-gene-in-escherichia-coli-pseudomonas-aeruginosa-and-acinetobacter-baumannii
#12
Delphine Girlich, Rémy A Bonnin, Agnes Jousset, Thierry Naas
Objectives: KPC-producing pathogens exhibit variable carbapenem susceptibility levels, which is probably the result of the genetic environment of the bla KPC genes. Here we determined the transcriptional start sites (TSSs) and the expression of the bla KPC-2 gene in various genetic contexts and in different hosts ( Escherichia coli , Pseudomonas aeruginosa and Acinetobacter baumannii ). Methods: The bla KPC-2 genes along with the upstream sequences derived from Tn 4401b (structure A), Tn 4401b interrupted by Tn 3 /IS 26 (structure B) and Tn 4401b interrupted by Tn 5563 (structure C) were cloned in two E...
March 5, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/28333270/environmental-and-genetic-modulation-of-the-phenotypic-expression-of-antibiotic-resistance
#13
Diarmaid Hughes, Dan I Andersson
Antibiotic resistance can be acquired by mutation or horizontal transfer of a resistance gene, and generally an acquired mechanism results in a predictable increase in phenotypic resistance. However, recent findings suggest that the environment and/or the genetic context can modify the phenotypic expression of specific resistance genes/mutations. An important implication from these findings is that a given genotype does not always result in the expected phenotype. This dissociation of genotype and phenotype has important consequences for clinical bacteriology and for our ability to predict resistance phenotypes from genetics and DNA sequences...
March 8, 2017: FEMS Microbiology Reviews
https://www.readbyqxmd.com/read/28333198/a-tetracycline-inducible-integrative-expression-system-for-streptococcus-pneumoniae
#14
Marina Meiers, Anke Laux, Daniela Eichinger, Anne Sexauer, Patrick Marx, Ralph Bertram, Reinhold Brückner
Inducible gene expression systems are very useful to analyze cellular processes. The ability to switch the expression state of genes of interest may even be crucial if essential traits or genetic instability are involved. An integrative plasmid, pTEX2, was designed using the (anhydro)tetracycline-inducible promoter Pxyl/tet from staphylococcal plasmid pRAB11 to control gene expression in Streptococcus pneumoniae. The system was evaluated by expressing genes of the two-component regulatory system ciaRH of S...
February 21, 2017: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/28333184/analysis-of-blashv-12-carrying-escherichia-coli-clones-and-plasmids-from-human-animal-and-food-sources
#15
Carla Andrea Alonso, Geovana Brenner Michael, Jun Li, Sergio Somalo, Carmen Simón, Yang Wang, Heike Kaspar, Kristina Kadlec, Carmen Torres, Stefan Schwarz
Objectives: This study aimed at characterizing 23 Escherichia coli isolates from various sources and their respective bla SHV-12 -carrying plasmids and sequencing one of these plasmids completely. Methods: Isolates were typed by XbaI-PFGE, MLST and PCR-based phylotyping. Transformed bla SHV-12 -carrying plasmids were examined by replicon typing, S1-nuclease, conjugation, EcoRI-HindIII-BamHI digests and plasmid MLST. Co-located resistance genes and integrons as well as the bla SHV-12 genetic environment were analysed by PCR and sequencing...
February 17, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/28332730/multicenter-validation-of-a-simplified-method-for-paroxysmal-nocturnal-hemoglobinuria-screening
#16
Arianna Gatti, Luigi Del Vecchio, Massimo Geuna, Matteo G Della Porta, Bruno Brando
According to the most credited and validated international guidelines, testing peripheral blood for the diagnosis of PNH is best accomplished with four color, two tubes or five to six color, single tube flow cytometric (FCM) procedures, with the demonstration of the deficiency of at least two GPI-linked surface proteins in two different cell subsets to corroborate the diagnosis (1-7). Each one of these state-of-the-art techniques ensure a high degree of sensitivity (i.e. down to 0.01% PNH granulocyte or red cell clones) with a high level of standardization and good reproducibility...
March 23, 2017: European Journal of Haematology
https://www.readbyqxmd.com/read/28332627/cross-neutralizing-anti-hiv-1-human-single-chain-variable-fragments-scfvs-against-cd4-binding-site-and-n332-glycan-identified-from-a-recombinant-phage-library
#17
Lubina Khan, Rajesh Kumar, Ramachandran Thiruvengadam, Hilal Ahmad Parray, Muzamil Ashraf Makhdoomi, Sanjeev Kumar, Heena Aggarwal, Madhav Mohata, Abdul Wahid Hussain, Raksha Das, Raghavan Varadarajan, Jayanta Bhattacharya, Madhu Vajpayee, K G Murugavel, Suniti Solomon, Subrata Sinha, Kalpana Luthra
More than 50% of HIV-1 infection globally is caused by subtype_C viruses. Majority of the broadly neutralizing antibodies (bnAbs) targeting HIV-1 have been isolated from non-subtype_C infected donors. Mapping the epitope specificities of bnAbs provides useful information for vaccine design. Recombinant antibody technology enables generation of a large repertoire of monoclonals with diverse specificities. We constructed a phage recombinant single chain variable fragment (scFv) library with a diversity of 7.8 × 10(8) clones, using a novel strategy of pooling peripheral blood mononuclear cells (PBMCs) of six select HIV-1 chronically infected Indian donors whose plasma antibodies exhibited potent cross neutralization efficiency...
March 23, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28332574/natural-variation-in-ctb4a-enhances-rice-adaptation-to-cold-habitats
#18
Zhanying Zhang, Jinjie Li, Yinghua Pan, Jilong Li, Lei Zhou, Hongli Shi, Yawen Zeng, Haifeng Guo, Shuming Yang, Weiwei Zheng, Jianping Yu, Xingming Sun, Gangling Li, Yanglin Ding, Liang Ma, Shiquan Shen, Luyuan Dai, Hongliang Zhang, Shuhua Yang, Yan Guo, Zichao Li
Low temperature is a major factor limiting rice productivity and geographical distribution. Improved cold tolerance and expanded cultivation to high-altitude or high-latitude regions would help meet growing rice demand. Here we explored a QTL for cold tolerance and cloned the gene, CTB4a (cold tolerance at booting stage), encoding a conserved leucine-rich repeat receptor-like kinase. We show that different CTB4a alleles confer distinct levels of cold tolerance and selection for variation in the CTB4a promoter region has occurred on the basis of environmental temperature...
March 23, 2017: Nature Communications
https://www.readbyqxmd.com/read/28332049/molecular-and-pharmacological-mechanisms-of-drug-resistance-an-evolving-paradigm
#19
Benedetta Colmegna, Lavinia Morosi, Maurizio D'Incalci
The high heterogeneity and genomic instability of malignant tumors explains why even responsive tumors contain cell clones that are resistant for many possible mechanisms involving intracellular drug inactivation, low uptake or high efflux of anticancer drugs from cancer cells, qualitative or quantitative changes in the drug target. Many tumors, however, are resistant because of insufficient exposure to anticancer drugs, due to pharmacokinetic reasons and inefficient and heterogeneous tumor drug distribution, related to a deficient vascularization and high interstitial pressure...
March 23, 2017: Handbook of Experimental Pharmacology
https://www.readbyqxmd.com/read/28332030/molecular-cloning-expression-and-characterization-of-pectin-methylesterase-ctpme-from-clostridium-thermocellum
#20
Vikky Rajulapati, Arun Goyal
Many phytopathogenic micro-organisms such as bacteria and fungi produce pectin methylesterases (PME) during plant invasion. Plants and insects also produce PME to degrade plant cell wall. In the present study, a thermostable pectin methylesterase (CtPME) from Clostridium thermocellum belonging to family 8 carbohydrate esterase (CE8) was cloned, expressed and purified. The amino acid sequence of CtPME exhibited similarity with pectin methylesterase from Erwinia chrysanthemi with 38% identity. The gene encoding CtPME was cloned into pET28a(+) vector and expressed using Escherichia coli BL21(DE3) cells...
March 22, 2017: Molecular Biotechnology
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