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https://www.readbyqxmd.com/read/29783967/characterization-of-a-novel-thermostable-and-xylose-tolerant-gh-39-%C3%AE-xylosidase-from-dictyoglomus-thermophilum
#1
Qi Li, Tao Wu, Zhipeng Qi, Linguo Zhao, Jianjun Pei, Feng Tang
BACKGROUND: β-D-xylosidase is a vital exoglycosidase with the ability to hydrolyze xylooligosaccharides to xylose and to biotransform some saponins by cleaving outer β-xylose. β-D-xylosidase is widely used as one of the xylanolytic enzymes in a diverse range of applications, such as fuel, food and the pharmaceutical industry; therefore, more and more studies have focused on the thermostable and xylose-tolerant β-D-xylosidases. RESULTS: A thermostable β-xylosidase gene (xln-DT) of 1509 bp was cloned from Dictyoglomus thermophilum and expressed in E...
May 21, 2018: BMC Biotechnology
https://www.readbyqxmd.com/read/29783112/the-neuropeptides-of-asian-freshwater-clam-corbicula-fluminea-as-new-molecular-biomarker-basing-on-the-responses-of-organophosphate-chemicals-exposure
#2
Qi Wang, Xiangsheng Hong, Huihui Chen, Lilai Yuan, Jinmiao Zha
In the present study, to discover new biomarker of Asian freshwater clam (Corbicula fluminea) to assess impact of environmental pollutions, cholecystokinin (CCK), conopressin, and Neuropeptide FF (FFamide) in C. fluminea were selected as potent biomarkers. Therefore, their full-length cDNAs were cloned and characterized to investigate the molecular characteristics and expression patterns of neuropeptides in C. fluminea. According to the sequence analysis, CCK, conopressin, and FFamide encoded proteins of 173, 152, and 90 amino acids, respectively...
May 18, 2018: Ecotoxicology and Environmental Safety
https://www.readbyqxmd.com/read/29781609/enhancement-and-analysis-of-human-anti-aflatoxin-b1-afb1-scfv-antibody-ligand-interaction-using-chain-shuffling
#3
Kuntalee Rangnoi, Kiattawee Choowongkomon, Richard O'Kennedy, Florian Rüker, Montarop Yamabhai
A human anti-aflatoxin B1 (AFB1) scFv antibody (yAFB1-c3), selected from a naïve human phage-displayed scFv library, was used as a template for improving and analysis of antibody-ligand interactions using the chain-shuffling technique. The variable heavy and variable light (VH/VL) shuffled library was constructed from the VH of 25 pre-selected clones recombined with the VL of yAFB1-c3 and vice versa. Affinity selection from these library demonstrated that the VH domain played an important role in the binding of scFv to free AFB1...
May 21, 2018: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/29781496/a-universal-platform-for-selection-and-high-resolution-phenotypic-screening-of-bacterial-mutants-using-the-nanowell-slide
#4
H Antypas, M Veses-Garcia, E Weibull, H Andersson-Svahn, A Richter-Dahlfors
The Petri dish and microtiter plate are the golden standard for selection and screening of bacteria in microbiological research. To improve on the limited resolution and throughput of these methods, we developed a universal, user-friendly platform for selection and high-resolution phenotypic screening based on the nanowell slide. This miniaturized platform has an optimal ratio between throughput and assay complexity, holding 672 nanowells of 500 nl each. As monoclonality is essential in bacterial genetics, we used FACS to inoculate each nanowell with a single bacterium in 15 min...
May 21, 2018: Lab on a Chip
https://www.readbyqxmd.com/read/29776931/shuffling-the-neutral-drift-of-unspecific-peroxygenase-in-saccharomyces-cerevisiae
#5
Javier Martin-Diaz, Carmen Paret, Eva García-Ruiz, Patricia Molina-Espeja, Miguel Alcalde
Unspecific peroxygenase (UPO) is a highly promiscuous biocatalyst and its selective mono(per)oxygenase activity makes it useful for many synthetic chemistry applications. Among the broad repertory of library creation methods for directed enzyme evolution, genetic drift allows neutral mutations to be accumulated gradually within a polymorphic network of variants. In this study, we conducted a campaign of genetic drift with UPO in Saccharomyces cerevisiae so that neutral mutations were simply added and recombined in vivo With low mutational loading and an activity threshold of 45% of the parent's native function, mutant libraries enriched in folded and active UPO variants were generated...
May 18, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29776924/tools-for-rapid-genetic-engineering-of-vibrio-fischeri
#6
Karen L Visick, Kelsey M Hodge-Hanson, Alice H Tischler, Allison K Bennett, Vincent Mastrodomenico
Vibrio fischeri is used as a model for number of processes, including symbiosis, quorum sensing, bioluminescence, and biofilm formation. Many of these studies depend on generating deletion mutants and complementing them. Engineering such strains, however, is a time-consuming, multi-step process that relies on cloning and subcloning. Here, we describe a set of tools that can be used to rapidly engineer deletions and insertions in the V. fischeri chromosome without cloning. We developed a uniform approach for generating deletions using PCR SOEing (Splicing by Overlap Extension) with antibiotic cassettes flanked by standardized linker sequences...
May 18, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29774030/changes-in-the-tcr%C3%AE-repertoire-and-tumor-immune-signature-from-a-cutaneous-melanoma-patient-immunized-with-the-csf-470-vaccine-a-case-report
#7
Mariana Aris, Alicia Inés Bravo, María Betina Pampena, Paula Alejandra Blanco, Ibel Carri, Daniel Koile, Patricio Yankilevich, Estrella Mariel Levy, María Marcela Barrio, José Mordoh
The allogeneic therapeutic vaccine CSF-470 has demonstrated a significant benefit over medium-dose IFNα2b in the distant metastasis-free survival for stages IIB-IIC-III cutaneous melanoma patients in a randomized phase II/III clinical trial (CASVAC-0401, NCT01729663). At the end of the 2-year CSF-470 immunization protocol, patient #006 developed several lung and one subcutaneous melanoma metastases; this later was excised. In this report, we analyzed the changes throughout vaccination of immune populations in blood and in the tumor tissue, with special focus on the T-cell repertoire...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29773895/crispr-cas9-system-targeting-regulatory-genes-of-hiv-1-inhibits-viral-replication-in-infected-t-cell-cultures
#8
Youdiil Ophinni, Mari Inoue, Tomohiro Kotaki, Masanori Kameoka
The CRISPR/Cas9 system provides a novel and promising tool for editing the HIV-1 proviral genome. We designed RNA-guided CRISPR/Cas9 targeting the HIV-1 regulatory genes tat and rev with guide RNAs (gRNA) selected from each gene based on CRISPR specificity and sequence conservation across six major HIV-1 subtypes. Each gRNA was cloned into lentiCRISPRv2 before co-transfection to create a lentiviral vector and transduction into target cells. CRISPR/Cas9 transduction into 293 T and HeLa cells stably expressing Tat and Rev proteins successfully abolished the expression of each protein relative to that in non-transduced and gRNA-absent vector-transduced cells...
May 17, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29773649/ptap-motif-duplication-in-the-p6-gag-protein-confers-a-replication-advantage-on-hiv-1-subtype-c
#9
Shilpee Sharma, Prabhu S Arunachalam, Malini Menon, Viswanath Ragupathy, Ravi Vijaya Satya, Joshua Jebaraj, Shambhu Ganeshappa Aralaguppe, Chaitra Rao, Sreshtha Pal, Shanmugam Saravanan, Kailapuri G Murugavel, Pachamuthu Balakrishnan, Suniti Solomon, Indira Hewlett, Udaykumar Ranga
HIV-1 subtype C (HIV-1C) may duplicate longer amino acid stretches in the p6 Gag protein, leading to the creation of an additional Pro-Thr/Ser-Ala-Pro (PTAP) motif necessary for viral packaging. However, the biological significance of a duplication of the PTAP motif for HIV-1 replication and pathogenesis has not been experimentally validated. In a longitudinal study of two different clinical cohorts of select HIV-1 seropositive, drug-naive individuals from India, we found that 8 of 50 of these individuals harbored a mixed infection of viral strains discordant for the PTAP duplication...
May 17, 2018: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/29771412/lncrna-cche1-in-the-proliferation-and-apoptosis-of-gastric-cancer-cells
#10
G Xu, Y Zhang, N Li, J-B Zhang, R Xu
OBJECTIVE: We aimed at investigating the effects of long non-coding RNA (lncRNA) CCHE1 on proliferation and apoptosis of gastric cancer cells. MATERIALS AND METHODS: qRT-PCR method was used to detect lncRNA CCHE1 in cancer cell lines. Cells with relative expression were selected to change the expression by transfecting with corresponding lentiviral vector, and nonsense transfection group and blank control group were also constructed; cell counting kit-8 (CCK-8) method was used to assess cell proliferation activity; colony formation assay was used to evaluate the rate of cell cloning; flow cytometry assay was used to determine the apoptosis rate and cell cycle...
May 2018: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/29771192/fine-scale-population-genetic-structure-and-within-tree-distribution-of-mating-types-of-venturia-effusa-cause-of-pecan-scab-in-the-u-s-a
#11
Clive Bock, Carolyn Young, Katherine Stevenson, Nikki Charlton
Scab (caused by Venturia effusa) is the major disease of pecan in the southeastern U.S.A. There is no information available on the fine scale population genetic diversity, or the occurrence of clonal types at small spatial scales that provides insight into inoculum sources and dispersal mechanisms, and potential opportunity for sexual reproduction. To investigate fine scale genetic diversity, 4 trees (cv. Wichita = populations) were sampled hierarchically: within each tree canopy, 4 approximately evenly spaced terminals (subpopulations) were selected and up to 6 leaflets (sub-subpopulations) were sampled from different compound leaves on each terminal...
May 17, 2018: Phytopathology
https://www.readbyqxmd.com/read/29770550/improved-robustness-of-an-ethanologenic-yeast-strain-through-adaptive-evolution-in-acetic-acid-is-associated-with-its-enzymatic-antioxidant-ability
#12
N Gurdo, G F Novelli Poisson, Á B Juárez, M C Ríos de Molina, M A Galvagno
AIMS: To investigate multiple tolerance of Saccharomyces cerevisiae obtained through a laboratory strategy of adaptive evolution in acetic acid, its relation with enzymatic ROS detoxification and bioethanol 2G production. METHODS AND RESULTS: After adaptive evolution in acetic acid, a clone (Y8A) was selected for its tolerance to high acetic acid concentrations (13 g l-1 ) in batch cultures. Y8A was resistant to multiple stresses: osmotic, thermic, oxidative, saline, ethanol, organic acid, phenolic compounds and slow freeze-thawing cycles...
May 16, 2018: Journal of Applied Microbiology
https://www.readbyqxmd.com/read/29769360/multiple-large-inversions-and-breakpoint-rewiring-of-gene-expression-in-the-evolution-of-the-fire-ant-social-supergene
#13
Yu-Ching Huang, Viet Dai Dang, Ni-Chen Chang, John Wang
Supergenes consist of co-adapted loci that segregate together and are associated with adaptive traits. In the fire ant Solenopsis invicta , two 'social' supergene variants regulate differences in colony queen number and other traits. Suppressed recombination in this system is maintained, in part, by a greater than 9 Mb inversion, but the supergene is larger. Has the supergene in S. invicta undergone multiple large inversions? The initial gene content of the inverted allele of a supergene would be the same as that of the wild-type allele...
May 16, 2018: Proceedings. Biological Sciences
https://www.readbyqxmd.com/read/29766518/pls-ipls-ga-pls-models-for-soluble-solids-content-ph-and-acidity-determination-in-intact-dovyalis-fruit-using-near-infrared-spectroscopy
#14
Mateus Wagner Dantas de Assis, Deborah Oliveira De Fusco, Rosangela Câmara Costa, Kássio Michell Gomes de Lima, Luis Carlos Cunha Júnior, Gustavo Henrique de Almeida Teixeira
BACKGROUND: Dovyalis species Dovyalis abyssinica Warb. and Dovyalis hebecarpa Warb. were introduced in Brazil, but the fruit quality of these species are not appropriate for fresh consumption due to the high titratable acidity (TA) and low soluble solids content (SSC). With the selection of new D. abyssinica clones with lower acidity and the hybridization of these two dovyalis specie (D. abyssinica and D. hebecarpa) the fruit quality improved and the better physical-chemical characteristics make them more suitable for fresh consumption...
May 16, 2018: Journal of the Science of Food and Agriculture
https://www.readbyqxmd.com/read/29766417/the-fto-gene-is-associated-with-growth-and-omega-3-6-ratio-in-asian-seabass
#15
Fei Sun, Rongjian Tu, Jun Hong Xia, Xiao Jun Liu, Gen Hua Yue
Polymorphisms in the FTO gene are associated with obesity and body mass index in humans and livestock. Little information of whether FTO plays an important role in aquaculture fish species is available. We cloned and characterized the FTO gene in an economically important food fish species: Asian seabass (Lates calcarifer). The full-length cDNA of the gene is 3679 bp, containing an ORF of 1935 bp encoding 644 amino acids, a 216 bp 5' UTR and a 1538 bp 3' UTR. The gene consisted of nine exons and eight introns and was 117,679 bp in length...
May 16, 2018: Marine Biotechnology
https://www.readbyqxmd.com/read/29765810/construction-of-a-synthetic-araneus-ventricosus-dragline-silk-gene-multimer-and-its-expression-in-escherichia-coli
#16
Tingting Liu, Anwen Liang, Ziqiang Liang, Guanghong Li, Fanghai Wang
One of the most representative core gene sequence of Araneus ventricosus dragline silk protein partial cDNA monomer (JN857964.2) was selected and multimerized using a "head-to-tail" strategy by compatible but nonregenerable sites at both ends resulting in a concatemer of 16 contiguous monomers. This concatemer was cloned into pET-28a(+) expression vector and transformed into Escherichia coli . A 52.6 kDa silk protein was successfully expressed and detected by SDS-PAGE and confirmed by Western blotting...
May 2018: 3 Biotech
https://www.readbyqxmd.com/read/29765375/functional-alleles-of-chicken-bg-genes-members-of-the-butyrophilin-gene-family-in-peripheral-t-cells
#17
Lei Chen, Michaela Fakiola, Karen Staines, Colin Butter, Jim Kaufman
γδ T cells recognize a wide variety of ligands in mammals, among them members of the butyrophilin (BTN) family. Nothing is known about γδ T cell ligands in chickens, despite there being many such cells in blood and lymphoid tissues, as well as in mucosal surfaces. The major histocompatibility complex (MHC) of chickens was discovered because of polymorphic BG genes, part of the BTN family. All but two BG genes are located in the BG region, oriented head-to-tail so that unequal crossing-over has led to copy number variation (CNV) as well as hybrid (chimeric) genes, making it difficult to identify true alleles...
2018: Frontiers in Immunology
https://www.readbyqxmd.com/read/29762716/crispor-intuitive-guide-selection-for-crispr-cas9-genome-editing-experiments-and-screens
#18
Jean-Paul Concordet, Maximilian Haeussler
CRISPOR.org is a web tool for genome editing experiments with the CRISPR-Cas9 system. It finds guide RNAs in an input sequence and ranks them according to different scores that evaluate potential off-targets in the genome of interest and predict on-target activity. The list of genomes is continuously expanded, with more 150 genomes added in the last two years. CRISPOR tries to provide a comprehensive solution from selection, cloning and expression of guide RNA as well as providing primers needed for testing guide activity and potential off-targets...
May 14, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29759152/minimal-residual-disease-detection-by-flow-cytometry-in-multiple-myeloma-why-and-how
#19
REVIEW
Mikhail Roshal
The outlook for myeloma patients has steadily improved with the introduction of newer drug combinations in recent years. Unlike older therapies that largely achieved only modest levels of neoplastic clone reduction, the newer drug combinations have led to deeper suppression of myeloma clones in most patients. Frequently the neoplastic clones become undetectable with traditional disease evaluation approaches. Recent studies using ultrasensitive disease monitoring have demonstrated that patients with disease undetectable by traditional techniques show wide heterogeneity in disease levels varying by several orders of magnitude...
January 2018: Seminars in Hematology
https://www.readbyqxmd.com/read/29757589/the-effect-of-differentially-designed-fusion-proteins-to-elicit-efficient-anti-human-thyroid-stimulating-hormone-immune-responses
#20
Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol-Karim Sheikhi, Mehdi Hedayati, Hossein Ghaderi-Zefrehi, Milad Alasvand
The production of human thyroid stimulating hormone (hTSH) immunoassays requires specific antibodies against hTSH which is a cumbersome process. Therefore, producing specific polyclonal antibodies against engineered recombinant fusion hTSH antigens would be of great significance. The best immunogenic region of the hTSH was selected based on in silico analyses and equipped with two different fusions. Standard methods were used for protein expression, purification, verification, structural evaluation, and immunizations of the white New Zealand rabbits...
April 2018: Iranian Journal of Allergy, Asthma, and Immunology
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