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https://www.readbyqxmd.com/read/28922761/rapid-mapping-and-cloning-of-the-virescent-1-gene-in-cotton-by-bulked-segregant-analysis-next-generation-sequencing-and-virus-induced-gene-silencing-strategies
#1
Jiankun Zhu, Jiedan Chen, Fengkai Gao, Chenyu Xu, Huaitong Wu, Kun Chen, Zhanfeng Si, Hu Yan, Tianzhen Zhang
Map-based gene cloning is a vital strategy for the identification of the quantitative trait loci or genes underlying important agronomic traits. The conventional map-based cloning method is powerful but generally time-consuming and labor-intensive. In this context, we introduce an improved bulked segregant analysis method in combination with a virus-induced gene silencing (VIGS) strategy for rapid and reliable gene mapping, identification and functional verification. This method was applied to a multiple recessive marker line of upland cotton, Texas 582 (T582), and identified unique genomic positions harboring mutant loci, showing the reliability and efficacy of this method...
July 10, 2017: Journal of Experimental Botany
https://www.readbyqxmd.com/read/28922567/determination-of-pd-l1-expression-in-effusions-from-mesothelioma-by-immuno-cytochemical-staining
#2
Mohammed S I Mansour, Tomas Seidal, Ulrich Mager, Amir Baigi, Katalin Dobra, Annika Dejmek
BACKGROUND: Malignant mesothelioma (MM) is an aggressive, fatal tumor. Current therapeutic options only marginally improve survival. Programmed cell death ligand 1 (PD-L1) is a dominant mediator of immunosuppression, binding to programmed cell death 1 (PD-1). PD-L1 is up-regulated in cancer cells, and the PD-1/PD-L1 pathway plays a critical role in tumor immune evasion, thus providing a target for antitumor therapy. Further, a correlation between PD-L1 expression and prognosis has been reported...
September 18, 2017: Cancer
https://www.readbyqxmd.com/read/28922396/biochemical-and-immunological-characterization-of-a-novel-monoclonal-antibody-against-mouse-leukotriene-b4-receptor-1
#3
Fumiyuki Sasaki, Tomoaki Koga, Kazuko Saeki, Toshiaki Okuno, Saiko Kazuno, Tsutomu Fujimura, Yasuyuki Ohkawa, Takehiko Yokomizo
Leukotriene B4 (LTB4) receptor 1 (BLT1) is a G protein-coupled receptor expressed in various leukocyte subsets; however, the precise expression of mouse BLT1 (mBLT1) has not been reported because a mBLT1 monoclonal antibody (mAb) has not been available. In this study, we present the successful establishment of a hybridoma cell line (clone 7A8) that produces a high-affinity mAb for mBLT1 by direct immunization of BLT1-deficient mice with mBLT1-overexpressing cells. The specificity of clone 7A8 was confirmed using mBLT1-overexpressing cells and mouse peripheral blood leukocytes that endogenously express BLT1...
2017: PloS One
https://www.readbyqxmd.com/read/28922060/risk-factors-for-nasal-colonization-by-methicillin-resistant-staphylococci-in-healthy-humans-in-professional-daily-contact-with-companion-animals-in-portugal
#4
Ana Catarina Rodrigues, Adriana Belas, Cátia Marques, Luís Cruz, Luís T Gama, Constança Pomba
Methicillin-resistant staphylococci (MRS), namely Staphylococcus aureus (MRSA) and Staphylococcus pseudintermedius (MRSP), are opportunistic agents of great importance in human and veterinary medicine. The aims of this study were to investigate the frequency, persistence, and risk factors associated with nasal colonization by MRS in people in daily contact with animals in Portugal. Seventy-nine out of 129 (61.2%) participants were found to be colonized by, at least, one methicillin-resistant (MR) staphylococci species (MR Staphylococcus epidermidis [n = 68], MRSA [n = 19], MR Staphylococcus haemolyticus [n = 7], MRSP [n = 2], and other coagulase-negative staphylococci [n = 4])...
September 18, 2017: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
https://www.readbyqxmd.com/read/28921690/tissue-specific-and-time-dependent-clonal-expansion-of-enu-induced-mutant-cells-in-gpt-delta-mice
#5
Takafumi Nakayama, Tomoko Sawai, Ikuko Masuda, Shinya Kaneko, Kazumi Yamauchi, Benjamin J Blyth, Yoshiya Shimada, Akira Tachibana, Shizuko Kakinuma
DNA mutations play a crucial role in the origins of cancer, and the clonal expansion of mutant cells is one of the fundamental steps in multistage carcinogenesis. In this study, we correlated tumor incidence in B6C3F1 mice during the period after exposure to N-ethyl-N-nitrosourea (ENU) with the persistence of ENU-induced mutant clones in transgenic gpt delta B6C3F1 mice. The induced gpt mutations afforded no selective advantage in the mouse cells and could be distinguished by a mutational spectrum that is characteristic of ENU treatment...
September 16, 2017: Environmental and Molecular Mutagenesis
https://www.readbyqxmd.com/read/28920565/regulatory-functions-of-trehalose-6-phosphate-synthase-in-the-chitin-biosynthesis-pathway-in-tribolium-castaneum-coleoptera-tenebrionidae-revealed-by-rna-interference
#6
Q W Chen, S Jin, L Zhang, Q D Shen, P Wei, Z M Wei, S G Wang, B Tang
RNA interference (RNAi) is a very effective technique for studying gene function and may be an efficient method for controlling pests. Trehalose-6-phosphate synthase (TPS), which plays a key role in the synthesis of trehalose and insect development, was cloned in Tribolium castaneum (Herbst) (TcTPS) and the putative functions were studied using RNAi via the injection of double-stranded RNA (dsRNA) corresponding to conserved TPS and trehalose-6-phosphate phosphatase domains. Expression analyses show that TcTPS is expressed higher in the fat body, while quantitative real-time polymerase chain reaction results show that the expression of four trehalase isoforms was significantly suppressed by dsTPS injection...
September 18, 2017: Bulletin of Entomological Research
https://www.readbyqxmd.com/read/28920412/molecular-cloning-purification-expression-and-characterization-of-%C3%AE-1-4-endoglucanase-gene-cel5a-from-eubacterium-cellulosolvens-sp-isolated-from-holstein-steer-s-rumen
#7
Tansol Park, Seongwon Seo, Teaksoon Shin, Byung-Wook Cho, Seongkeun Cho, Byeongwoo Kim, Jong K Ha, Jakyeom Seo
Objective: This study was conducted to isolate the cellulolytic microorganism from a rumen of Holstein steer and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, the rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 99% similarity with Eubacterium cellulosolvens Ce2 (Accession number: AY178842.1). The Cel5A from isolated E. cellulosolvens sp...
September 18, 2017: Asian-Australasian Journal of Animal Sciences
https://www.readbyqxmd.com/read/28920397/-effects-of-interleukin-34-expressed-by-human-bone-marrow-derived-mesenchymal-stem-cells-on-thp-1-cells
#8
Xueming Gao, Shun Zhang, Zhaojun Xu, Yan Shan, Zihong Wang, Ting Cai, Xue Jiang
To construct recombinant eukaryotic expression plasmid vector of human IL-34 gene, and to study the effects of IL-34 expressed by human bone marrow-derived mesenchymal stem cells (hBM-MSCs) on THP-1 cells. Full-length IL-34 encoding sequence was amplified by PCR. And this fragment was cloned into the plasmid pIRES2-EGFP. Western blotting and ELISA were used to analyze the expression of IL-34 in hBM-MSCs. THP-1 cells were cultured with hBM-MSCs medium containing IL-34 protein. Real-time PCR detected the effects of IL-34 on the expression of IL-10 and TNFα in THP-1 cells...
April 25, 2017: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
https://www.readbyqxmd.com/read/28920005/the-frequency-of-neoantigens-per-somatic-mutation-rather-than-overall-mutational-load-or-number-of-predicted-neoantigens-per-se-is-a-prognostic-factor-in-ovarian-clear-cell-carcinoma
#9
Hirokazu Matsushita, Kosei Hasegawa, Katsutoshi Oda, Shogo Yamamoto, Akira Nishijima, Yuichi Imai, Kayo Asada, Yuji Ikeda, Takahiro Karasaki, Keiichi Fujiwara, Hiroyuki Aburatani, Kazuhiro Kakimi
Neoantigens derived from tumor-specific somatic mutations are excellent targets for anti-tumor immune responses. In ovarian clear cell carcinoma (OCCC), checkpoint blockade yields durable responses in a subset of patients. To approach the question of why only some patients respond, we first investigated neoantigen loads and immune signatures using exome sequencing and expression array data for 74 OCCC patients treated conventionally. Neither the number of missense mutations nor total predicted neoantigens assessed in the tumor correlated with clinical outcomes...
2017: Oncoimmunology
https://www.readbyqxmd.com/read/28919972/cytogenetic-maps-of-homoeologous-chromosomes-a-h01-and-d-h01-and-their-integration-with-the-genome-assembly-in-gossypium-hirsutum
#10
Yuling Liu, Zhen Liu, Renhai Peng, Yuhong Wang, Zhongli Zhou, Xiaoyan Cai, Xingxing Wang, Zhenmei Zhang, Kunbo Wang, Fang Liu
Cytogenetic maps of Gossypium hirsutum (Linnaeus, 1753) homoeologous chromosomes Ah01 and Dh01 were constructed by fluorescence in situ hybridization (FISH), using eleven homoeologous-chromosomes-shared bacterial artificial chromosomes (BACs) clones and one chromosome-specific BAC clone respectively. We compared the cytogenetic maps with the genetic linkage and draft genome assembly maps based on a standardized map unit, relative map position (RMP), which allowed a global view of the relationship of genetic and physical distances along each chromosome, and assembly quality of the draft genome assembly map...
2017: Comparative Cytogenetics
https://www.readbyqxmd.com/read/28919547/deep-sequencing-to-resolve-complex-diversity-of-apicomplexan-parasites-in-platypuses-and-echidnas-proof-of-principle-for-wildlife-disease-investigation
#11
Jan Šlapeta, Stefan Saverimuttu, Larry Vogelnest, Cheryl Sangster, Frances Hulst, Karrie Rose, Paul Thompson, Richard Whittington
The short-beaked echidna (Tachyglossus aculeatus) and the platypus (Ornithorhynchus anatinus) are iconic egg-laying monotremes (Mammalia: Monotremata) from Australasia. The aim of this study was to demonstrate the utility of diversity profiles in disease investigations of monotremes. Using small subunit (18S) rDNA amplicon deep-sequencing we demonstrated the presence of apicomplexan parasites and confirmed by direct and cloned amplicon gene sequencing Theileria ornithorhynchi, Theileria tachyglossi, Eimeria echidnae and Cryptosporidium fayeri...
September 12, 2017: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/28919531/a-thermostable-alkaliphilic-protein-disulfide-isomerase-from-bacillus-subtilis-dr8806-cloning-expression-biochemical-characterization-and-molecular-dynamics-simulation
#12
Milad Lagzian, Ali Shahraki, Mahdiyeh Besharatian, Ahmad Asoodeh
Disulfide bonds are among the most important factors related to correct folding of the proteins. Protein disulfide isomerase (PDI) is the enzyme responsible for the correct formation and isomerization of these bonds. It is rarely studied so far and none of them showed industrial properties. In this study, the gene encoding for a putative PDI from Bacillus subtilis DR8806 was identified, cloned and expressed in Escherichia coli. It was encoded a 23.26kDa protein. The enzyme was purified by GST affinity chromatography with a specific activity of 1227u/mg...
September 14, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28919436/the-notch-pathway-regulates-the-second-mitotic-wave-cell-cycle-independently-of-bhlh-proteins
#13
Abhishek Bhattacharya, Ke Li, Manon Quiquand, Gerard Rimesso, Nicholas E Baker
Notch regulates both neurogenesis and cell cycle activity to coordinate precursor cell generation in the differentiating Drosophila eye. Mosaic analysis with mitotic clones mutant for Notch components was used to identify the pathway of Notch signaling that regulates the cell cycle in the Second Mitotic Wave. Although S phase entry depends on Notch signaling and on the transcription factor Su(H), the transcriptional co-activator Mam and the bHLH repressor genes of the E(spl)-Complex were not essential, although these are Su(H) coactivators and targets during the regulation of neurogenesis...
September 14, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28919361/molecular-characterisation-ontogeny-and-expression-analysis-of-melanoma-differentiation-associated-factor-5-mda5-from-asian-seabass-lates-calcarifer
#14
Anutosh Paria, M Makesh, Aparna Chaudhari, C S Purushothaman, K V Rajendran
MDA5 is the pivotal member of the retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) and is reported to play a crucial role in type I IFN-mediated responses against pathogen-associated molecular patterns (PAMPs), especially nucleic acids. In this study, we have identified and cloned the full-length cDNA sequence of MDA5, which comprises 3398 nucleotides and encodes for a putative protein of 978 AA length, in Asian seabass, Lates calcarifer. From the putative amino acid sequence of AsMDA5, four different conserved domains could be predicted: two N-terminal CARD domains, a DExDc domain, a HELICc domain and a C-terminal RIG-1_C-RD domain...
September 15, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28919331/production-and-characterization-of-monoclonal-antibodies-against-cathepsin-b-and-cathepsin-b-like-proteins-of-naegleria-fowleri
#15
Gi-Sang Seong, Hae-Jin Sohn, Heekyoung Kang, Ga-Eun Seo, Jong-Hyun Kim, Ho-Joon Shin
Naegleria fowleri causes fatal primary amoebic meningoencephalitis (PAM) in humans and experimental animals. In previous studies, cathepsin B (nfcpb) and cathepsin B-like (nfcpb-L) genes of N. fowleri were cloned, and it was suggested that refolding rNfCPB and rNfCPB-L proteins could play important roles in host tissue invasion, immune response evasion and nutrient uptake. In this study, we produced anti-NfCPB and anti-NfCPB-L monoclonal antibodies (McAb) using a cell fusion technique, and observed their immunological characteristics...
September 14, 2017: Experimental Parasitology
https://www.readbyqxmd.com/read/28919326/characterization-of-a-pkr-inhibitor-from-the-pathogenic-ranavirus-ambystoma-tigrinum-virus-using-a-heterologous-vaccinia-virus-system
#16
Trung P Huynh, James K Jancovich, Latha Tripuraneni, Michael C Heck, Jeffrey O Langland, Bertram L Jacobs
Ambystoma tigrinum virus (ATV) (family Iridoviridae, genus Ranavirus) was isolated from diseased tiger salamanders (Ambystoma tigrinum stebbinsi) from the San Rafael Valley in southern Arizona, USA in 1996. Genomic sequencing of ATV, as well as other members of the genus, identified an open reading frame that has homology to the eukaryotic translation initiation factor, eIF2α (ATV eIF2α homologue, vIF2αH). Therefore, we asked if the ATV vIF2αH could also inhibit PKR. To test this hypothesis, the ATV vIF2αH was cloned into vaccinia virus (VACV) in place of the well-characterized VACV PKR inhibitor, E3L...
September 14, 2017: Virology
https://www.readbyqxmd.com/read/28919197/imported-poultry-meat-as-a-source-of-extended-spectrum-cephalosporin-resistant-cmy-2-producing-salmonella-heidelberg-and-s-minnesota-in-european-union-2014-2015
#17
Joana Campos, Joana Mourão, Leonor Silveira, Margarida Saraiva, Cristina Belo Correia, Ana Paula Maçãs, Luísa Peixe, Patrícia Antunes
Extended-spectrum cephalosporin (ESC)-resistant Salmonella has been described at low level in EU, nevertheless the increasing importation of poultry meat could be an important source of epidemic strains carrying ESC-resistant genes. We evaluated ESC resistance and characterized genetic platforms as well as the clonal relatedness of Salmonella isolates from poultry meat products imported into Portugal. All Salmonella isolates recovered from samples of fresh meat destined to be imported into the EU in the scope of Portuguese official border control between 2014 and 2015 were studied...
September 14, 2017: International Journal of Antimicrobial Agents
https://www.readbyqxmd.com/read/28919178/the-presence-of-abalone-egg-laying-hormone-like-peptide-in-the-central-nervous-system-and-ovary-of-the-spotted-babylon-babylonia-areolata
#18
Jirawat Saetan, Piyakorn Boonyoung, Uraporn Vongvatcharanon, Thanapong Kruangkum, Kanjana Khornchatri, Pinij Thaweethamsewee, Prasert Sobhon, Prapee Sretarugsa
Recently, the neuronal classification of the ivory shell Spotted Babylon, Babylonia areolata, was readily demonstrated. Regarding its importance as marine economic molluscan species, the attempt to understand the neuroendocrine regulation is necessary. This study firstly demonstrated the neurosecretory cells as well as the existence and distribution of the egg-laying hormone (ELH)-like peptide in the central nervous system (CNS) and ovary of the B. areolata. The neurosecretory cell was characterized by the cytoplasmic purple dot-like structure as stained by the Gomori's paraldehyde fuchsin...
September 14, 2017: Acta Histochemica
https://www.readbyqxmd.com/read/28918746/escherichia-coli-signal-peptidase-recognizes-and-cleaves-archaeal-signal-sequence
#19
Majida Atta Muhammad, Samia Falak, Naeem Rashid, Qurra-Tul-Ann Afza Gardner, Nasir Ahmad, Tadayuki Imanaka, Muhammad Akhtar
Tk1884, an open reading frame encoding α-amylase in Thermococcus kodakarensis, was cloned with the native signal sequence and expressed in Escherichia coli. Heterologous gene expression resulted in secretion of the recombinant protein to the extracellular culture medium. Extracellular α-amylase activity gradually increased after induction. Tk1884 was purified from the extracellular medium, and its molecular mass determined by electrospray ionization mass spectrometry indicated the cleavage of a few amino acids...
July 2017: Biochemistry. Biokhimii︠a︡
https://www.readbyqxmd.com/read/28918565/forward-genetic-screens-identify-a-role-for-the-mitochondrial-her2-in-e-2-hexenal-responsiveness
#20
Alessandra Scala, Rossana Mirabella, Joachim Goedhart, Michel de Vries, Michel A Haring, Robert C Schuurink
This work adds a new player, HER2, downstream of the perception of E-2-hexenal, a green leaf volatile, and shows that E-2-hexenal specifically changes the redox status of the mitochondria. It is widely accepted that plants produce and respond to green leaf volatiles (GLVs), but the molecular components involved in transducing their perception are largely unknown. The GLV E-2-hexenal inhibits root elongation in seedlings and, using this phenotype, we isolated E-2-hexenal response (her) Arabidopsis thaliana mutants...
September 16, 2017: Plant Molecular Biology
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