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https://www.readbyqxmd.com/read/28442397/molecular-cloning-characterization-and-expression-analysis-of-heat-shock-protein-90-in-albino-northern-snakehead-channa-argus
#1
Aiguo Zhou, Shaolin Xie, Zhenlu Wang, Muhammad Junaid, Lanfen Fan, Chao Wang, Qiao Ye, Yanfeng Chen, De-Sheng Pei, Jixing Zou
The great albino northern snakehead Channa argus is habitual to only the Sichuan Jialing Rivers System in China, making its introduction difficult to other riverine systems. Here, we characterized heat shock protein 90 (AcaHSP90) and probed its molecular responses toward the environmental stressors that C. argus can face during its introduction and breeding in the other southern latitudes of China. To serve the purpose, cDNA encoding of AcaHSP90 were cloned and characterized in albino C. argus. The cDNA was 2752bps that contained an open reading frame (ORF), encoding a 726-amino-acid polypeptide of 83...
April 22, 2017: Gene
https://www.readbyqxmd.com/read/28439822/improved-denaturation-of-small-rna-duplexes-and-its-application-for-northern-blotting
#2
C Jake Harris, David C Baulcombe, Attila Molnar
Small RNAs (sRNAs) are short (18-30 nucleotide) noncoding RNA molecules, which control gene expression and pathogen response in eukaryotes. They are associated with and guide nucleases to target nucleic acids by nucleotide base pairing. We found that current techniques for small RNA detection are adversely affected by the presence of complementary RNA. Thus we established FDF-PAGE (fully denaturing formaldehyde polyacrylamide gel electrophoresis), which dramatically improves denaturation efficiency and subsequently the detection of sequestered sRNAs...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28412374/a-novel-mitochondrial-atp6-frameshift-mutation-causing-isolated-complex-v-deficiency-ataxia-and-encephalomyopathy
#3
Christopher Benjamin Jackson, Dagmar Hahn, Barbara Schröter, Uwe Richter, Brendan Battersby, Thomas Schmitt-Mechelke, Paula Martinen, Jean-Marc Nuoffer, André Schaller
We describe a novel frameshift mutation in the mitochondrial ATP6 gene in a 4-year-old girl associated with ataxia, microcephaly, developmental delay and intellectual disability. A heteroplasmic frameshift mutation in the MT-ATP6 gene was confirmed in the patient's skeletal muscle and blood. The mutation was not detectable in the mother's DNA extracted from blood or buccal cells. Enzymatic and oxymetric analysis of the mitochondrial respiratory system in the patients' skeletal muscle and skin fibroblasts demonstrated an isolated complex V deficiency...
April 12, 2017: European Journal of Medical Genetics
https://www.readbyqxmd.com/read/28401641/a-conserved-%C3%AE-proteobacterial-small-rna-contributes-to-osmoadaptation-and-symbiotic-efficiency-of-rhizobia-on-legume-roots
#4
Marta Robledo, Alexandra Peregrina, Vicenta Millán, Natalia I García-Tomsig, Omar Torres-Quesada, Pedro F Mateos, Anke Becker, José I Jiménez-Zurdo
Small non-coding RNAs (sRNAs) are expected to have pivotal roles in the adaptive responses underlying symbiosis of nitrogen-fixing rhizobia with legumes. Here, we provide primary insights into the function and activity mechanism of the Sinorhizobium meliloti trans-sRNA NfeR1 (Nodule Formation Efficiency RNA). Northern blot probing and transcription tracking with fluorescent promoter-reporter fusions unveiled high nfeR1 expression in response to salt stress and throughout the symbiotic interaction. The strength and differential regulation of nfeR1 transcription are conferred by a motif, which is conserved in nfeR1 promoter regions in α-proteobacteria...
April 12, 2017: Environmental Microbiology
https://www.readbyqxmd.com/read/28400331/four-paralog-gelsolin-genes-are-differentially-expressed-in-the-earthworm-lumbricus-terrestris
#5
Prasath Thiruketheeswaran, Paul Thomalla, Evelyn Krüger, Horst Hinssen, Jochen D'Haese
We have identified and characterized four distinct variants of the gelsolin-related protein (EWAM P1-P4) in the earthworm L. terrestris. All of these proteins biochemically qualify as gelsolins since they sever actin filaments in a calcium dependent manner. P1, P2 and P3 are present in the Lumbricus body wall muscle whereas in the gizzard muscle P3 and P4 were found. P1-P4 are encoded by four paralog genes and are differentially expressed in various muscle cell tissues. While the genes for P1 and P2 contain one intron, there was no intron in both P3 and P4 genes...
April 8, 2017: Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
https://www.readbyqxmd.com/read/28396683/preparation-and-neutralization-efficacy-of-igy-antibodies-raised-against-deinagkistrodon-acutus-venom
#6
Jinhua Liu, Qiyi He, Wenwen Wang, Bin Zhou, Bo Li, Yingfeng Zhang, Cong Luo, Diancheng Chen, Jia Tang, Xiaodong Yu
BACKGROUND: The five-paced pit viper (Deinagkistrodon acutus), endemic to China and northern Vietnam, is responsible for most snakebites in the Chinese territory. Antivenom produced from horses is the main treatment for snakebites, but it may cause numerous clinical side effects and have other disadvantages involved in their production such as the welfare of animals. The present study was conducted aiming to develop an alternative antibody (IgY) from the egg yolk of leghorn chickens immunized with snake venom...
2017: Journal of Venomous Animals and Toxins Including Tropical Diseases
https://www.readbyqxmd.com/read/28396653/reduced-hepatic-carcinoembryonic-antigen-related-cell-adhesion-molecule-1-level-in-obesity
#7
Garrett Heinrich, Harrison T Muturi, Khadijeh Rezaei, Qusai Y Al-Share, Anthony M DeAngelis, Thomas A Bowman, Hilda E Ghadieh, Simona S Ghanem, Deqiang Zhang, Robert S Garofalo, Lei Yin, Sonia M Najjar
Impairment of insulin clearance is being increasingly recognized as a critical step in the development of insulin resistance and metabolic disease. The carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) promotes insulin clearance. Null deletion or liver-specific inactivation of Ceacam1 in mice causes a defect in insulin clearance, insulin resistance, steatohepatitis, and visceral obesity. Immunohistological analysis revealed reduction of hepatic CEACAM1 in obese subjects with fatty liver disease...
2017: Frontiers in Endocrinology
https://www.readbyqxmd.com/read/28385057/a-comprehensive-molecular-investigation-of-%C3%AE-thalassemia-in-an-iranian-cohort-from-different-provinces-of-north-iran
#8
Hajar Eftekhari, Ahmad Tamaddoni, Hassan Mahmoudi Nesheli, Mohsen Vakili, Sadegh Sedaghat, Ali Banihashemi, Mandana Azizi, Reza Youssefi Kamangar, Haleh Akhavan-Niaki
α-Thalassemia (α-thal) is the most common monogenic disease that is caused by the absence or reduced expression of α-globin genes. The aim of this study was to investigate common α-globin mutations and their associated haplotypes in four northern provinces of Iran (Gilan, Mazandaran, Golestan, Khorasan). One thousand, one hundred and ninety-one persons were tested for α-thal mutations by gap-polymerase chain reaction (PCR), reverse dot-blot hybridization, restriction fragment length polymorphism (RFLP) analysis and sequencing...
April 7, 2017: Hemoglobin
https://www.readbyqxmd.com/read/28375845/effect-of-adrenocorticotropic-hormone-on-ucp1-gene-expression-in-brown-adipocytes
#9
Hirendra M Biswas
BACKGROUND: Like other tissues, adrenocorticotropic hormone (ACTH) can produce its effect on brown adipose tissue (BAT). This study was taken to understand the direct effect of ACTH action on thermogenin gene expression and possible relation with α receptors and caffeine with this hormone. METHODS: Brown fat precursor cells were isolated from interscapular BAT of young mice and grown in culture. The cells were exposed to norepinephrine (NE) and other agents. Total RNA was isolated after harvesting the cells, and northern blot analysis was performed...
April 4, 2017: Journal of Basic and Clinical Physiology and Pharmacology
https://www.readbyqxmd.com/read/28373490/schizosaccharomyces-pombe-polysome-profile-analysis-and-rna-purification
#10
Dieter A Wolf, Jürg Bähler, Jo Ann Wise
Polysome profile analysis is widely used by investigators studying the mechanism and regulation of translation. The method described here uses high-velocity centrifugation of whole cell extracts on linear sucrose gradients to separate 40S and 60S ribosomal subunits from 80S monosomes and polysomes. Cycloheximide is included in the lysis buffer to "freeze" polysomes by blocking translation. After centrifugation, the gradient is fractionated and RNA (and/or protein) is prepared from each fraction for subsequent analysis of individual species using northern or western blots...
April 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28370207/micrornas-based-imaging-techniques-in-cancer-diagnosis-and-therapy
#11
Maryam Keshavarzi, Mohammad Jafar Rezaie, Saba Sorayayi, Mohsen Mohammadi, Amir Ghaderi, Ayoob Rostamzadeh, Aria Masoudifar, Hamed Mirzaei
Cancer is one of the most serious global health concerns in different populations. Several studies indicated that there are many potentially promising cellular and molecular targets for cancer therapy within cancer cells and their microenvironment. Among different cellular and molecular targets involved in cancer pathogenesis, microRNAs (miRNAs) are well known as key targets for cancer therapy. miRNAs are one of main classes of non-coding RNAs. These molecules play important roles in different critical processes of cancer pathogenesis...
March 29, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28368073/molecular-analysis-of-the-ramra-locus-in-clinical-klebsiella-pneumoniae-isolates-with-reduced-susceptibility-to-tigecycline
#12
Cristina Belmar Campos, Martin Aepfelbacher, Moritz Hentschke
Mutations in ramR, a negative regulator of ramA which stimulates transcription of acrA/-B encoding the multidrug efflux pump AcrAB-TolC, were recently shown to result in reduced susceptibility to tigecycline in Klebsiella pneumoniae. We analysed six non-duplicate K. pneumoniae isolates with elevated MICs to tigecycline. All isolates showed transcriptional up-regulation of ramA and acrB as demonstrated by Northern blot and quantitative real-time PCR. Sequencing of the ramR gene revealed deletions in five of the isolates and a premature stop codon in one isolate...
April 3, 2017: New Microbiologica
https://www.readbyqxmd.com/read/28363873/production-of-recombinant-human-acid-%C3%AE-glucosidase-with-high-mannose-glycans-in-gnt1-rice-for-the-treatment-of-pompe-disease
#13
Jae-Wan Jung, Nguyen-Xuan Huy, Hyo-Boon Kim, Nan-Sun Kim, Do Van Giap, Moon-Sik Yang
Lysosomal storage diseases are a group of inherited metabolic disorders. Patients are treated with enzyme replacement therapy (ERT), in which the replacement enzymes are required to carry terminal mannose or mannose 6-phosphate residues to allow efficient uptake into target cells and tissues. N-acetylglucosaminyltransferase-I (GnTI) mediates N-glycosylation in the cis cisternae of the Golgi apparatus by adding N-acetylglucosamine to the exposed terminal mannose residue of core N-glycan structures for further processing...
March 29, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28361900/micrornas-associated-with-caste-determination-and-differentiation-in-a-primitively-eusocial-insect
#14
David H Collins, Irina Mohorianu, Matthew Beckers, Vincent Moulton, Tamas Dalmay, Andrew F G Bourke
In eusocial Hymenoptera (ants, bees and wasps), queen and worker adult castes typically arise via environmental influences. A fundamental challenge is to understand how a single genome can thereby produce alternative phenotypes. A powerful approach is to compare the molecular basis of caste determination and differentiation along the evolutionary trajectory between primitively and advanced eusocial species, which have, respectively, relatively undifferentiated and strongly differentiated adult castes. In the advanced eusocial honeybee, Apis mellifera, studies suggest that microRNAs (miRNAs) play an important role in the molecular basis of caste determination and differentiation...
March 31, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28337962/linc00052-promotes-gastric-cancer-cell-proliferation-and-metastasis-via-activating-wnt-%C3%AE-catenin-signaling-pathway
#15
Yuqiang Shan, Rongchao Ying, Zhong Jia, Wencheng Kong, Yi Wu, Sixin Zheng, Huicheng Jin
Gastric cancer (GC) is one of the most common malignant tumors of digestive system. The etiology of GC is complex and much more attention should be paid on the genetic factors. In this study, we explored the role and function of LINC00052 in GC. We applied qRT-PCR and northern blot to detect the expression of LINC00052 and found it was highly expressed during GC. We also investigated the effects of LINC00052 on tumor prognosis and progression, and found LINC00052 indicated poor prognosis and tumor progression...
March 23, 2017: Oncology Research
https://www.readbyqxmd.com/read/28334889/the-primary-transcriptome-of-neisseria-meningitidis-and-its-interaction-with-the-rna-chaperone-hfq
#16
Nadja Heidrich, Saskia Bauriedl, Lars Barquist, Lei Li, Christoph Schoen, Jörg Vogel
Neisseria meningitidis is a human commensal that can also cause life-threatening meningitis and septicemia. Despite growing evidence for RNA-based regulation in meningococci, their transcriptome structure and output of regulatory small RNAs (sRNAs) are incompletely understood. Using dRNA-seq, we have mapped at single-nucleotide resolution the primary transcriptome of N. meningitidis strain 8013. Annotation of 1625 transcriptional start sites defines transcription units for most protein-coding genes but also reveals a paucity of classical σ70-type promoters, suggesting the existence of activators that compensate for the lack of -35 consensus sequences in N...
March 17, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/28330156/effect-of-salicylic-acid-on-expression-level-of-genes-related-with-isoprenoid-pathway-in-centella-centella-asiatica-l-urban-cells
#17
Nguyen Hoang Loc, Nguyen Thanh Giang, Nguyen Duc Huy
In this study, we report the expression level of CaSQS, CabAS and CaCYS, the genes involved in phytosterol and triterpene metabolic pathway of centella (Centella asiatica (L.) Urban), in cells elicited with salicylic acid (50-200 µM). Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis indicated CaSQS, CabAS, and CaCYS genes expressed in both the wild-type and cultured cells (with and without elicitation). In elicited cells, expressions of CaSQS, CabAS, and CaCYS genes showed strong dependence on salicylic acid concentration and elicitation day...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28317896/use-of-rna-seq-data-to-identify-and-validate-rt-qpcr-reference-genes-for-studying-the-tomato-pseudomonas-pathosystem
#18
Marina A Pombo, Yi Zheng, Zhangjun Fei, Gregory B Martin, Hernan G Rosli
The agronomical relevant tomato-Pseudomonas syringae pv. tomato pathosystem is widely used to explore and understand the underlying mechanisms of the plant immune response. Transcript abundance estimation, mainly through reverse transcription-quantitative PCR (RT-qPCR), is a common approach employed to investigate the possible role of a candidate gene in certain biological process under study. The accuracy of this technique relies heavily on the selection of adequate reference genes. Initially, genes derived from other techniques (such as Northern blots) were used as reference genes in RT-qPCR experiments, but recent studies in different systems suggest that many of these genes are not stably expressed...
March 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28296577/genome-wide-identification-and-characterization-of-small-rnas-in-rhodobacter-capsulatus-and-identification-of-small-rnas-affected-by-loss-of-the-response-regulator-ctra
#19
Marc P Grüll, Lourdes Peña-Castillo, Martin E Mulligan, Andrew S Lang
Small non-coding RNAs (sRNAs) are involved in the control of numerous cellular processes through various regulatory mechanisms, and in the past decade many studies have identified sRNAs in a multitude of bacterial species using RNA sequencing (RNA-seq). Here, we present the first genome-wide analysis of sRNA sequencing data in Rhodobacter capsulatus, a purple nonsulfur photosynthetic alphaproteobacterium. Using a recently developed bioinformatics approach, sRNA-Detect, we detected 422 putative sRNAs from R...
March 15, 2017: RNA Biology
https://www.readbyqxmd.com/read/28286035/identification-and-characterization-of-two-putative-micrornas-encoded-by-bombyx-mori-cypovirus
#20
Zhong-Hua Pan, Ping Wu, Kun Gao, Cheng-Xiang Hou, Guang-Xing Qin, Tao Geng, Xi-Jie Guo
Viral microRNAs (miRNAs) have been demonstrated to play important roles in virus-host interactions. Some RNA virus-encoded miRNAs have been reported to promote viral replication and may be used as potential drug targets. Bombyx mori cypovirus (BmCPV), an important pathogen of silkworm, is a double-stranded RNA virus frequently causing serious damages in sericulture. Research on miRNA encoded by BmCPV may be useful to elucidate the BmCPV-host interaction and to develop new anti-viral methods. In our previous study, small RNA libraries of the midgut of BmCPV-infected silkworm have been generated by deep sequencing and several BmCPV-encoded putative miRNAs were predicted...
March 8, 2017: Virus Research
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