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https://www.readbyqxmd.com/read/29332216/characteristics-of-arg-carrying-plasmidome-in-the-cultivable-microbial-community-from-wastewater-treatment-system-under-high-oxytetracycline-concentration
#1
Yanhong Shi, Hong Zhang, Zhe Tian, Min Yang, Yu Zhang
Studies on antibiotic production wastewater have shown that even a single antibiotic can select for multidrug resistant bacteria in aquatic environments. It is speculated that plasmids are an important mechanism of multidrug resistance (MDR) under high concentrations of antibiotics. Herein, two metagenomic libraries were constructed with plasmid DNA extracted from cultivable microbial communities in a biological wastewater treatment reactor supplemented with 0 (CONTROL) or 25 mg/L of oxytetracycline (OTC-25)...
January 13, 2018: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29331528/expression-of-a-thermotolerant-laccase-from-pycnoporus-sanguineus-in-trichoderma-reesei-and-its-application-in-the-degradation-of-bisphenol-a
#2
Jie Zhao, Shenquan Zeng, Ying Xia, Liming Xia
The laccase gene from Pycnoporus sanguineus was cloned and inserted between the strong Pcbh1 promoter and the Tcbh1 terminator from Trichoderma reesei to form the recombinant plasmid pCH-lac. Using Agrobacterium-mediated technique, the pCH-lac was integrated into the chromosomes of T. reesei. Twenty positive transformants were obtained by employing hygromycin B as a selective agent. PCR was used to confirm that the laccase gene was integrated into the chromosomal DNA of T. reesei. Laccase production by recombinant transformants was performed in shaking flasks, and the activity of laccase reached 8...
January 10, 2018: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/29331527/construction-of-sake-yeast-with-low-production-of-dimethyl-trisulfide-precursor-by-a-self-cloning-method
#3
Yuriko Ikeda, Atsuko Isogai, Yuta Moriyoshi, Ryoko Kanda, Kazuhiro Iwashita, Tsutomu Fujii
Dimethyl trisulfide (DMTS) is the primary component responsible for "hineka", the stale aroma of Japanese sake. Deletion of the MRI1 or MDE1 gene of sake yeast, encoding 5'-methylthioribose-1-phosphate isomerase and 5'-methylthioribulose-1-phosphate dehydratase, respectively, has been reported to greatly reduce the amount of DMTS precursor (DMTS-P1) in sake and to suppress the formation of DMTS during storage. In this study, we constructed sake yeast strains lacking MRI1 gene function by a self-cloning method...
January 10, 2018: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/29331150/production-of-eicosapentaenoic-acid-by-application-of-a-delta-6-desaturase-with-the-highest-ala-catalytic-activity-in-algae
#4
Haisu Shi, Xue Luo, Rina Wu, Xiqing Yue
Dunaliella salina is a unicellular green alga with a high α-linolenic acid (ALA) level, but a low eicosapentaenoic acid (EPA) level. In a previous analysis of the catalytic activity of delta 6 fatty acid desaturase (FADS6) from various species, FADS6 from Thalassiosira pseudonana (TpFADS6), a marine diatom, showed the highest catalytic activity for ALA. In this study, to enhance EPA production in D. salina, FADS6 from D. salina (DsFADS6) was identified, and substrate specificities for DsFADS6 and TpFADS6 were characterized...
January 13, 2018: Microbial Cell Factories
https://www.readbyqxmd.com/read/29330804/in-vivo-manipulation-of-hif-1%C3%AE-expression-during-glioma-genesis
#5
Patricia D B Tiburcio, Séan B Lyne, L Eric Huang
Hypoxia has long been recognized as a driving force of tumor progression and therapeutic resistance, and the transcription factor HIF-1α is believed to play a crucial role in these processes. Here we describe an efficient RCAS/Nes-TVA system that allows for in vivo manipulation of HIF-1α expression in the mouse neural progenitor cells. Simple production of the recombinant avian virus RCAS enables quick delivery of gene of interest through injection into the neural progenitors of transgenic mice expressing the viral cognate receptor TVA under the nestin promoter...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29330557/linear-doggybone-dna-vaccine-induces-similar-immunological-responses-to-conventional-plasmid-dna-independently-of-immune-recognition-by-tlr9-in-a-pre-clinical-model
#6
Alex Allen, Chuan Wang, Lisa J Caproni, Gessa Sugiyarto, Elena Harden, Leon R Douglas, Patrick J Duriez, Kinga Karbowniczek, Jon Extance, Paul J Rothwell, Ifeayinwa Orefo, John P Tite, Freda K Stevenson, Christian H Ottensmeier, Natalia Savelyeva
Vaccination with DNA that encodes cancer antigens is a simple and convenient way to raise immunity against cancer and has already shown promise in the clinical setting. Conventional plasmid DNA is commonly used which together with the encoded antigen also includes bacterial immunostimulatory CpG motifs to target the DNA sensor Toll-like receptor 9. Recently DNA vaccines using doggybone DNA (dbDNA™), have been developed without the use of bacteria. The cell-free process relies on the use of Phi29 DNA polymerase to amplify the template followed by protelomerase TelN to complete individual closed linear DNA...
January 12, 2018: Cancer Immunology, Immunotherapy: CII
https://www.readbyqxmd.com/read/29330186/effect-of-plasmid-design-and-type-of-integration-event-on-recombinant-protein-expression-in-pichia-pastoris
#7
Thomas Vogl, Leigh Gebbie, Robin W Palfreyman, Robert Speight
Pichia pastoris (syn. Komagataella phaffii) is one of the most common eukaryotic expression systems for heterologous protein production. Expression cassettes are typically integrated in the genome to obtain stable expression strains. In contrast to Saccharomyces cerevisiae, where short overhangs are sufficient to target highly specific integration, long overhangs are more efficient in P. pastoris and ectopic integration of foreign DNA can occur. Here, we aimed to elucidate the influence of ectopic integration by high throughput screening of >700 transformants and whole genome sequencing of 27 transformants...
January 12, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29330182/fluorescence-recovery-allows-the-implementation-of-a-fluorescence-reporter-gene-platform-applicable-for-the-detection-and-quantification-of-horizontal-gene-transfer-in-anoxic-environments
#8
Rafael Pinilla, Leise Riber, Søren J Sørensen
The study of horizontal gene transfer (HGT) in microbial communities has been revolutionized by significant advances in cultivation-independent methods based on fluorescence reporter gene technologies. Recently, the combination of these novel approaches with flow cytometry has presented itself as one of the most powerful tools to study the spread of mobile genetic elements (MGEs) in the environment. However, the use of fluorescent markers like green fluorescent protein (GFP) and mCherry is limited by environmental constraints that affect the correct maturation of their fluorophores, such as oxygen availability and pH levels...
January 12, 2018: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/29329863/preparation-of-gentiooligosaccharides-using-trichoderma-viride-%C3%AE-glucosidase
#9
Fei Wang, Jing Wu, Sheng Chen
The recombinant plasmid pPIC9K-bgl1 containing β-glucosidase bgl1 from Trichoderma viride was constructed by overlapping PCR and integrated into Pichia pastoris KM71. In order to assist the formation of disulfide bonds and thus improve protein folding efficiency, protein disulfide isomerase pdi was co-expressed in the P. pastoris KM71/pPIC9K-bgl1/pPICZ-A-pdi strain, and fermentation in flasks resulted in enzyme activity of 143 U/ml. The enzyme activity of β-glucosidase reached 1402 U/ml following optimisation of fermentation conditions in a 3...
May 15, 2018: Food Chemistry
https://www.readbyqxmd.com/read/29329720/discoveries-of-extrachromosomal-circles-of-dna-in-normal-and-tumor-cells
#10
REVIEW
Teressa Paulsen, Pankaj Kumar, M Murat Koseoglu, Anindya Dutta
While the vast majority of cellular DNA in eukaryotes is contained in long linear strands in chromosomes, we have long recognized some exceptions like mitochondrial DNA, plasmids in yeasts, and double minutes (DMs) in cancer cells where the DNA is present in extrachromosomal circles. In addition, specialized extrachromosomal circles of DNA (eccDNA) have been noted to arise from repetitive genomic sequences like telomeric DNA or rDNA. Recently eccDNA arising from unique (nonrepetitive) DNA have been discovered in normal and malignant cells, raising interesting questions about their biogenesis, function and clinical utility...
January 9, 2018: Trends in Genetics: TIG
https://www.readbyqxmd.com/read/29329377/endometrial-stem-cell-derived-g-csf-attenuates-endometrial-fibrosis-via-sonic-hedgehog-transcriptional-activator-gli2
#11
Xiaona Lin, Yanling Zhang, Yibin Pan, Shilin He, Yongdong Dai, Bingqing Zhu, Cheng Wei, Liaobing Xin, Wenzhi Xu, Chunsheng Xiang, Songying Zhang
Intrauterine adhesion (IUA) is characterized by endometrial fibrosis, which ultimately leads to menstrual abnormalities, infertility and recurrent miscarriages. The Shh/Gli2 pathway plays a critical role in tissue fibrogenesis and regeneration; Gli2 activation induces profibrogenic effects in various tissues, such as the liver and kidney. However, the role of Gli2 in endometrial fibrosis remains unknown. The purpose of this study was to test the hypothesis that activated Gli2 promotes endometrial fibrosis. Endometrial samples from moderate and severe IUA patients exhibited significantly enhanced expression of Gli2 compared with normal endometrial samples and mild IUA samples...
January 10, 2018: Biology of Reproduction
https://www.readbyqxmd.com/read/29328481/ttf1%C3%A2-np-induces-protective-autophagy-during-apoptosis-by-inhibiting-the-akt-mtor-pathway-and-activating-jnk-in-human-liver-cancer-cells
#12
Xuan Zhang, Meilan Zhang, Bin Xiao, Wenjun Jiao, Silin Zhang, Xuewu Zhang
TTF1‑NP is a flavonoid nanoparticle based on 5,2',4'‑trihydroxy‑6,7,5'‑trimethoxyflavone (TTF1), which is derived from the medicinal plant Sorbaria sorbifolia that grows in the Changbai Mountain. We previously demonstrated antitumor effects of TTF1‑NP in human hepatoma including induction of apoptosis and inhibition of angiogenesis, migration and invasion. Herein, we examined the effects of TTF1‑NP on autophagy and its relationship with apoptosis, and explored potential underlying mechanisms in human hepatoma cell lines...
January 5, 2018: Oncology Reports
https://www.readbyqxmd.com/read/29328435/mir%C3%A2-214-reduces-cisplatin-resistance-by-targeting-netrin%C3%A2-1-in-bladder-cancer-cells
#13
Jiao Liu, Jianbin Bi, Zeliang Li, Zhenhua Li, Xiankui Liu, Chuize Kong
miR‑214 has been reported to be downregulated in several cancer types, such as bladder cancer. However, its involvement in apoptosis and chemoresistance has not been investigated. The present study aimed to clarify the biological function of miR‑214 and potential mechanisms in chemoresistance of bladder cancer cells. Reverse transcription‑quantitative polymerase chain reaction demonstrated that miR‑214 was downregulated in bladder cancer tissues compared with the level in normal tissues. miR‑214 was downregulated in bladder cancer cell lines compared with the level in the normal cell line SV‑HUC‑1...
January 10, 2018: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29328362/microrna-493-5p-promotes-apoptosis-and-suppresses-proliferation-and-invasion-in-liver-cancer-cells-by-targeting-vamp2
#14
Guannan Wang, Xiaosan Fang, Meng Han, Xiaoming Wang, Qiang Huang
The aim of the present study was to explore the role of miR‑493-5p in liver cancer tissues and cell lines, and its effect on cell behavioral characteristics. The expression of miR-493-5p was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) in liver cancer tissues and cell lines (hepatic cell line HL-7702 and the liver cancer cell lines HCCC-9810, HuH-7 and HepG2). In addition, the mechanism by which miR-493-5p mediates its effects was analyzed via the transfection of miR-493-5p mimic and negative control miRNA into HepG2 cells...
January 2, 2018: International Journal of Molecular Medicine
https://www.readbyqxmd.com/read/29327679/natural-and-artificial-strategies-to-control-the-conjugative-transmission-of-plasmids
#15
María Getino, Fernando de la Cruz
Conjugative plasmids are the main carriers of transmissible antibiotic resistance (AbR) genes. For that reason, strategies to control plasmid transmission have been proposed as potential solutions to prevent AbR dissemination. Natural mechanisms that bacteria employ as defense barriers against invading genomes, such as restriction-modification or CRISPR-Cas systems, could be exploited to control conjugation. Besides, conjugative plasmids themselves display mechanisms to minimize their associated burden or to compete with related or unrelated plasmids...
January 2018: Microbiology Spectrum
https://www.readbyqxmd.com/read/29327444/developing-a-flexible-high-efficiency-agrobacterium-mediated-sorghum-transformation-system-with-broad-application
#16
Ping Che, Ajith Anand, Emily Wu, Jeffry D Sander, Marissa K Simon, Weiwei Zhu, Amy L Sigmund, Gina Zastrow-Hayes, Michael Miller, Donglong Liu, Shai J Lawit, Zuo-Yu Zhao, Marc C Albertsen, Todd J Jones
Sorghum is the fifth most widely planted cereal crop in the world and is commonly cultivated in arid and semi-arid regions such as Africa. Despite its importance as a food source, sorghum genetic improvement through transgenic approaches has been limited because of an inefficient transformation system. Here we report a ternary vector (also known as co-habitating vector) system using a recently described pVIR accessory plasmid that facilitates efficient Agrobacterium-mediated transformation of sorghum. We report regeneration frequencies ranging from 6-29% in Tx430 using different selectable markers and single copy, backbone free "quality events" ranging from 45-66% of the total events produced...
January 11, 2018: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/29327301/lncrna-gas8-as1-inhibits-cell-proliferation-through-atg5-mediated-autophagy-in-papillary-thyroid-cancer
#17
Yuan Qin, Wei Sun, Hao Zhang, Ping Zhang, Zhihong Wang, Wenwu Dong, Liang He, Ting Zhang, Liang Shao, Wenqian Zhang, Changhao Wu
PURPOSE: The long non-coding RNA GAS8 antisense RNA 1 (lncRNA GAS8-AS1) is a tumor suppressor in papillary thyroid cancer (PTC), but the mechanisms underlying how GAS8-AS1 regulates PTC biology remain unclear. Here, we evaluated the molecular function of GAS8-AS1 in regulating autophagy in PTC cell lines. METHODS: GAS8-AS1 was overexpressed and knocked down in PTC cell lines by transfecting with expression plasmids or short interfering RNAs (siRNAs). Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8)...
January 11, 2018: Endocrine
https://www.readbyqxmd.com/read/29326210/complete-genome-sequencing-of-acinetobacter-sp-strain-logew2-3-isolated-from-the-pellet-of-a-white-stork-reveals-a-novel-class-d-beta-lactamase-gene
#18
Ulrike Blaschke, Evelyn Skiebe, Michael Kaatz, Paul G Higgins, Yvonne Pfeifer, Gottfried Wilharm
Whole-genome sequencing of Acinetobacter sp. strain LoGeW2-3, isolated from the pellet of a white stork (Ciconia ciconia), reveals the presence of a plasmid of 179,399 bp encoding a CRISPR-Cas (clustered regularly interspaced short palindromic repeats and associated genes) system of the I-F type, and the chromosomally encoded novel class D beta-lactamase OXA-568.
January 11, 2018: Genome Announcements
https://www.readbyqxmd.com/read/29325127/inhibiting-trophoblast-par-1-overexpression-suppresses-sflt-1-induced-anti-angiogenesis-and-abnormal-vascular-remodeling-a-possible-therapeutic-approach-for-preeclampsia
#19
Yin Zhao, YanFang Zheng, XiaoXia Liu, QingQing Luo, Di Wu, XiaoPing Liu, Li Zou
STUDY QUESTION: Is it possible to improve vascular remodeling by inhibiting the excessive expression of protease activated receptor 1 (PAR-1) in trophoblast of abnormal placenta? SUMMARY ANSWER: Inhibition of trophoblast PAR-1 overexpression may promote placental angiogenesis and vascular remodeling, offering an alternative therapeutic approach for preeclampsia. WHAT IS KNOWN ALREADY: PAR-1 is high-affinity receptor of thrombin. Thrombin increases sFlt-1 secretion in trophoblast via the activation of PAR-1...
January 9, 2018: Molecular Human Reproduction
https://www.readbyqxmd.com/read/29325123/packaging-of-dinoroseobacter-shibae-dna-into-gene-transfer-agent-particles-is-not-random
#20
Jürgen Tomasch, Hui Wang, April T K Hall, Diana Patzelt, Matthias Preusse, Jörn Petersen, Henner Brinkmann, Boyke Bunk, Sabin Bhuju, Michael Jarek, Robert Geffers, Andrew S Lang, Irene Wagner-Döbler
Gene transfer agents (GTAs) are phage-like particles which contain a fragment of genomic DNA of the bacterial or archaeal producer and deliver this to a recipient cell. GTA gene clusters are present in the genomes of almost all marine Rhodobacteraceae (Roseobacters) and might be important contributors to horizontal gene transfer (HGT) in the world's oceans. For all organisms studied so far, no obvious evidence of sequence specificity or other non-random process responsible for packaging genomic DNA into GTAs has been found...
January 9, 2018: Genome Biology and Evolution
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