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Molecular cloning

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https://www.readbyqxmd.com/read/28436337/molecular-cloning-expression-and-characterization-of-e2f-transcription-factor-4-from-antheraea-pernyi
#1
M N Abbas, S Kausar, Y-X Sun, Y Sun, L Wang, C Qian, G-Q Wei, B-J Zhu, C-L Liu
The E2F transcription factor family is distributed widely in eukaryotes and has been well studied among mammals. In the present study, the E2F transcription factor 4 (E2F4) gene was isolated from fat bodies of Antheraea pernyi and sequenced. E2F4 comprised a 795 bp open reading frame encoding a deduced amino acid sequence of 264 amino acid residues. The recombinant protein was expressed in Escherichia coli (Transetta DE3), and anti-E2F4 antibodies were prepared. The deduced amino acid sequence displayed significant homology to an E2F4-like protein from Bombyx mori L...
April 24, 2017: Bulletin of Entomological Research
https://www.readbyqxmd.com/read/28436098/lecrk-v-an-l-type-lectin-receptor-kinase-in-haynaldia-villosa-plays-positive-role-in-resistance-to-wheat-powdery-mildew
#2
Zongkuan Wang, Jiangyue Cheng, Anqi Fan, Jia Zhao, Zhongyu Yu, Yingbo Li, Heng Zhang, Jin Xiao, Faheem Mohammad, Haiyan Wang, Aizhong Cao, Liping Xing, Xiue Wang
Plant sense potential microbial pathogen by using pattern recognition receptors (PRRs) to recognize pathogen associated molecular patterns (PAMPs). The Lectin receptor like kinase genes (LecRKs) are involved in various cellular processes mediated by signal transduction pathways. In the present study, an L-type lectin receptor kinase gene LecRK-V was cloned from Haynaldia villosa, a diploid wheat relative which is highly resistant to powdery mildew. The expression of LecRK-V was rapidly up-regulated by Bgt inoculation and chitin treatment...
April 24, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28435199/genetic-characterization-of-the-non-structural-protein-3-gene-of-bluetongue-virus-serotype-2-isolate-from-india
#3
Raghavendra Sumanth Pudupakam, Shobana Raghunath, Meghanath Pudupakam, Sreenivasulu Daggupati
AIM: Sequence analysis and phylogenetic studies based on non-structural protein-3 (NS3) gene are important in understanding the evolution and epidemiology of bluetongue virus (BTV). This study was aimed at characterizing the NS3 gene sequence of Indian BTV serotype-2 (BTV2) to elucidate its genetic relationship to global BTV isolates. MATERIALS AND METHODS: The NS3 gene of BTV2 was amplified from infected BHK-21 cell cultures, cloned and subjected to sequence analysis...
March 2017: Veterinary World
https://www.readbyqxmd.com/read/28435108/molecular-characterization-and-antimicrobial-susceptibility-of-the-ca-mrsa-isolated-from-healthcare-workers-tehran-iran
#4
Solmaz Ohadian Moghadam, Mohammad Modoodi Yaghooti, Nasrin Pourramezan, Mohammad Reza Pourmand
INTRODUCTION: Methicillin resistant Staphylococcus aureus (MRSA) has become as a nosocomial pathogen worldwide. Considering the importance of MRSA typing for understanding the evolution and dissemination of these strains, we studied the molecular characteristics of MRSA colonized healthcare workers (HCWs). METHODOLOGY: All MRSA isolated from HCWs, were genotyped using staphylococcal cassette chromosome mec (SCCmec) with multiplex PCR assay, multilocus sequence typing (MLST) and spa typing...
April 20, 2017: Microbial Pathogenesis
https://www.readbyqxmd.com/read/28434792/cloning-and-expression-patterns-of-neuromedin-u-and-its-receptors-in-pigs
#5
Xiang Li, Mingfu Niu, Juan Su, Zhiyu Ma, Mengmeng Jin, Wenna Qiao, Ying Zhang, Yanyan Feng, Na An, Yuanlong Hou, Sheng Yang, Suo Chuan, Zhihai Lei
Neuromedin U (NMU) is a highly conserved neuropeptide that performs a variety of physiological functions in animals via neuromedin U receptor-1 (NMUR1) and neuromedin U receptor-2 (NMUR2). In this study, we cloned the pig NMU, NMUR1 and NMUR2 genes. Bioinformatics analysis demonstrated that the pig NMU cDNA encoded the amino acids Phe-Leu-Phe-Arg-Pro-Arg-Asn-NH2 at the C-terminus and that the NMU receptors, which are G-protein-coupled receptors (GPCRs), contained the seven transmembrane domains typical of GPCRs...
April 6, 2017: Neuropeptides
https://www.readbyqxmd.com/read/28434412/molecular-characterization-and-transcriptional-analysis-of-the-female-enriched-chondroitin-proteoglycan-2-of-toxocara-canis
#6
G X Ma, R Q Zhou, L Hu, Y L Luo, Y F Luo, H H Zhu
Toxocara canis is an important but neglected zoonotic parasite, and is the causative agent of human toxocariasis. Chondroitin proteoglycans are biological macromolecules, widely distributed in extracellular matrices, with a great diversity of functions in mammals. However, there is limited information regarding chondroitin proteoglycans in nematode parasites. In the present study, a female-enriched chondroitin proteoglycan 2 gene of T. canis (Tc-cpg-2) was cloned and characterized. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to measure the transcription levels of Tc-cpg-2 among tissues of male and female adult worms...
April 24, 2017: Journal of Helminthology
https://www.readbyqxmd.com/read/28434226/in-situ-microprobe-single-cell-capillary-electrophoresis-mass-spectrometry-metabolic-reorganization-in-single-differentiating-cells-in-the-live-vertebrate-x-laevis-embryo
#7
Rosemary M Onjiko, Erika P Portero, Sally A Moody, Peter Nemes
Knowledge of single-cell metabolism would provide a powerful window to cell activity changes as cells differentiate to all the tissues of the vertebrate embryo. However, single-cell mass spectrometry technologies have not yet been made compatible with complex three-dimensional changes and rapidly decreasing cell sizes during early development of the embryo. Here, we bridge this technological gap by integrating capillary microsampling, microscale metabolite extraction, and capillary electrophoresis electrospray ionization mass spectrometry (CE-ESI-MS) to enable direct metabolic metabolic analysis of identified cells in the live frog embryo (Xenopus laevis)...
April 22, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28433658/molecular-cloning-of-esr2-and-gene-expression-analysis-of-esr1-and-esr2-in-the-pituitary-gland-of-the-chinese-alligator-alligator-sinensis-during-female-reproductive-cycle
#8
Ruidong Zhang, Yanan Yin, Long Sun, Peng Yan, Yongkang Zhou, Rong Wu, Xiaobing Wu
Estrogens play critical roles in reproductive physiology via estrogen receptors (ESRs) in vertebrates, including reptiles. Chinese alligator (Alligator sinensis) is an endemic and endangered reptile species in China. In the present study, we cloned ESR2 gene from the ovary using rapid amplification of cDNA ends (RACE), investigated the spatial expression of ESRs in various tissues and temporal expression of ESRs in the pituitary glands during the reproductive cycle in Chinese alligators by quantitative real-time PCR (qPCR)...
April 19, 2017: Gene
https://www.readbyqxmd.com/read/28433252/nrasq61r-immunohistochemistry-detects-both-nrasq61r-and-krasq61r-mutations-in-colorectal-cancer
#9
Jie-Yang Jhuang, Chang-Tsu Yuan, Yu-Lin Lin, Mei-Ling Cheng, Jau-Yu Liau, Jia-Huei Tsai
The NRASQ61R monoclonal antibody (clone sp174) is a mutation-specific antibody that is increasingly being used to detect the NRAS(Q61R) mutation in melanomas. This antibody has been reported to be highly correlated with the NRAS(Q61R) mutation status in melanomas and follicular neoplasms of the thyroid gland. However, its utility in colorectal carcinoma (CRC) has remained largely unknown. In this study, we assessed the sensitivity, specificity, and diagnostic utility of NRASQ61R immunohistochemistry in a cohort consisting of tissue sections of 113 CRCs, which were molecularly profiled for the KRAS, NRAS, and BRAF mutations...
April 19, 2017: Pathology
https://www.readbyqxmd.com/read/28431894/molecular-cloning-and-functional-characterization-of-duck-nucleotide-binding-oligomerization-domain-1-nod1
#10
Huilin Li, Hui Jin, Yaqian Li, Dejian Liu, Mohamed Frahat Foda, Yunbo Jiang, Rui Luo
Nucleotide-binding oligomerization domain 1 (NOD1) is an imperative cytoplasmic pattern recognition receptor (PRR) and considered as a key member of the NOD-like receptor (NLR) family which plays a critical role in innate immunity through sensing microbial components derived from bacterial peptidoglycan. In the current study, the full-length of duck NOD1 (duNOD1) cDNA from duck embryo fibroblasts (DEFs) was cloned. Multiple sequence alignment and phylogenetic analysis demonstrated that duNOD1 exhibited a strong evolutionary relationship with chicken and rock pigeon NOD1...
April 19, 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/28431015/large-scale-collection-of-full-length-cdna-and-transcriptome-analysis-in-hevea-brasiliensis
#11
Yuko Makita, Kiaw Kiaw Ng, G Veera Singham, Mika Kawashima, Hideki Hirakawa, Shusei Sato, Ahmad Sofiman Othman, Minami Matsui
Natural rubber has unique physical properties that cannot be replaced by products from other latex-producing plants or petrochemically produced synthetic rubbers. Rubber from Hevea brasiliensis is the main commercial source for this natural rubber that has a cis-polyisoprene configuration. For sustainable production of enough rubber to meet demand elucidation of the molecular mechanisms involved in the production of latex is vital. To this end, we firstly constructed rubber full-length cDNA libraries of RRIM 600 cultivar and sequenced around 20,000 clones by the Sanger method and over 15,000 contigs by Illumina sequencer...
April 1, 2017: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes
https://www.readbyqxmd.com/read/28429853/molecular-cloning-of-mda5-phylogenetic-analysis-of-rig-i-like-receptors-rlrs-and-differential-gene-expression-of-rlrs-interferons-and-proinflammatory-cytokines-after-in%C3%A2-vitro-challenge-with-ipnv-isav-and-sav-in-the-salmonid-cell-line-to
#12
I-K G Nerbøvik, M A Solheim, H Ø Eggestøl, A Rønneseth, R A Jakobsen, H I Wergeland, G T Haugland
The RIG-I receptors RIG-I, MDA5 and LGP2 are involved in viral recognition, and they have different ligand specificity and recognize different viruses. Activation of RIG-I-like receptors (RLRs) leads to production of cytokines essential for antiviral immunity. In fish, most research has focused on interferons, and less is known about the production of proinflammatory cytokines during viral infections. In this study, we have cloned the full-length MDA5 sequence in Atlantic salmon, and compared it with RIG-I and LGP2...
April 21, 2017: Journal of Fish Diseases
https://www.readbyqxmd.com/read/28429795/cd200-positive-cancer-associated-fibroblasts-augment-the-sensitivity-of-epidermal-growth-factor-receptor-mutation-positive-lung-adenocarcinomas-to-egfr-tyrosine-kinase-inhibitors
#13
Masayuki Ishibashi, Shinya Neri, Hiroko Hashimoto, Tomoyuki Miyashita, Tatsuya Yoshida, Yuka Nakamura, Hibiki Udagawa, Keisuke Kirita, Shingo Matsumoto, Shigeki Umemura, Kiyotaka Yoh, Seiji Niho, Masahiro Tsuboi, Kenkichi Masutomi, Koichi Goto, Atsushi Ochiai, Genichiro Ishii
Cancer associated fibroblasts (CAFs) play important roles in the chemotherapeutic process, especially through influencing the resistance of tumor cells to molecular targeted therapy. Here we report the existence of a special subpopulation of patient-specific-CAFs that augment the sensitivity of EGFR gene mutation-positive lung cancer to the EGFR-tyrosine kinase inhibitor (EGFR-TKI), gefitinib. When cocultured with EGFR mutation positive lung cancer cells, these CAFs increased the apoptic effect of gefitinib on cancer cells, whereas, in the absence of gefitinib, they did not affect cancer cell viability...
April 21, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28429735/phylogenetic-analysis-of-metastatic-progression-in-breast-cancer-using-somatic-mutations-and-copy-number-aberrations
#14
David Brown, Dominiek Smeets, Borbála Székely, Denis Larsimont, A Marcell Szász, Pierre-Yves Adnet, Françoise Rothé, Ghizlane Rouas, Zsófia I Nagy, Zsófia Faragó, Anna-Mária Tőkés, Magdolna Dank, Gyöngyvér Szentmártoni, Nóra Udvarhelyi, Gabriele Zoppoli, Lajos Pusztai, Martine Piccart, Janina Kulka, Diether Lambrechts, Christos Sotiriou, Christine Desmedt
Several studies using genome-wide molecular techniques have reported various degrees of genetic heterogeneity between primary tumours and their distant metastases. However, it has been difficult to discern patterns of dissemination owing to the limited number of patients and available metastases. Here, we use phylogenetic techniques on data generated using whole-exome sequencing and copy number profiling of primary and multiple-matched metastatic tumours from ten autopsied patients to infer the evolutionary history of breast cancer progression...
April 20, 2017: Nature Communications
https://www.readbyqxmd.com/read/28428794/fine-mapping-of-a-novel-defective-glume-1-dg1-mutant-which-affects-vegetative-and-spikelet-development-in-rice
#15
Haiping Yu, Banpu Ruan, Zhongwei Wang, Deyong Ren, Yu Zhang, Yujia Leng, Dali Zeng, Jiang Hu, Guangheng Zhang, Li Zhu, Zhenyu Gao, Guang Chen, Longbiao Guo, Wenfu Chen, Qian Qian
In cereal crops, vegetative and spikelet development play important roles in grain yield and quality, but the genetic mechanisms that control vegetative and spikelet development remain poorly understood in rice. Here, we identified a new rice mutant, defective glume 1 (dg1) mutant from cultivar Zhonghua11 after ethyl methanesulfonate treatment. The dg1 mutant displayed the dwarfism with small, rolled leaves, which resulted from smaller cells and more bulliform cells. The dg1 mutant also had an enlarged leaf angle and defects in brassinosteroid signaling...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28428782/a-recombinant-potato-virus-y-infectious-clone-tagged-with-the-rosea1-visual-marker-pvy-ros1-facilitates-the-analysis-of-viral-infectivity-and-allows-the-production-of-large-amounts-of-anthocyanins-in-plants
#16
Teresa Cordero, Mohamed A Mohamed, Juan-José López-Moya, José-Antonio Daròs
Potato virus Y (PVY) is a major threat to the cultivation of potato and other solanaceous plants. By inserting a cDNA coding for the Antirrhinum majus Rosea1 transcription factor into a PVY infectious clone, we created a biotechnological tool (PVY-Ros1) that allows infection by this relevant plant virus to be tracked by the naked eye with no need for complex instrumentation. Rosea1 is an MYB-type transcription factor whose expression activates the biosynthesis of anthocyanin pigments in a dose-specific and cell-autonomous manner...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28428772/molecular-characterization-of-staphylococcus-aureus-isolated-from-bovine-mastitis-and-close-human-contacts-in-south-african-dairy-herds-genetic-diversity-and-inter-species-host-transmission
#17
Tracy Schmidt, Marleen M Kock, Marthie M Ehlers
Staphylococcus aureus is one of the most common etiological agents of contagious bovine mastitis worldwide. The purpose of this study was to genetically characterize a collection of S. aureus isolates (bovine = 146, human = 12) recovered from cases of bovine mastitis and nasal swabs of close human contacts in the dairy environment. Isolates were screened for a combination of clinically significant antimicrobial and virulence gene markers whilst the molecular epidemiology of these isolates and possible inter-species host transmission was investigated using a combination of genotyping techniques...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28427900/molecular-cloning-and-characterization-of-kiss1-in-brandt-s-voles-lasiopodomys-brandtii
#18
Yan Chen, Lan Liu, Zhengguang Li, Dawei Wang, Ning Li, Ying Song, Cong Guo, Xiaohui Liu
Kisspeptin, encoded by kiss1, has been regarded as a major modulator of mammalian puberty and fertility due to its stimulation on GnRH. Brandt's vole is one of the main pest species on the Inner Mongolian steppes for its striking reproductive capacity and kiss1 is a key candidate gene related to reproductive regulatory cascades. In this study, kiss1 cDNA was cloned from the hypothalamus of Brandt's voles and kiss1 mRNA levels were investigated in different tissues, and at different developmental stages, using high-throughput real-time PCR...
April 18, 2017: Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
https://www.readbyqxmd.com/read/28427478/morphological-and-molecular-characteristics-of-malayfilaria-sofiani-uni-mat-udin-takaoka-n-g-n-sp-nematoda-filarioidea-from-the-common-treeshrew-tupaia-glis-diard-duvaucel-mammalia-scandentia-in-peninsular-malaysia
#19
Shigehiko Uni, Ahmad Syihan Mat Udin, Takeshi Agatsuma, Weerachai Saijuntha, Kerstin Junker, Rosli Ramli, Hasmahzaiti Omar, Yvonne Ai-Lian Lim, Sinnadurai Sivanandam, Emilie Lefoulon, Coralie Martin, Daicus Martin Belabut, Saharul Kasim, Muhammad Rasul Abdullah Halim, Nur Afiqah Zainuri, Subha Bhassu, Masako Fukuda, Makoto Matsubayashi, Masashi Harada, Van Lun Low, Chee Dhang Chen, Narifumi Suganuma, Rosli Hashim, Hiroyuki Takaoka, Mohd Sofian Azirun
BACKGROUND: The filarial nematodes Wuchereria bancrofti (Cobbold, 1877), Brugia malayi (Brug, 1927) and B. timori Partono, Purnomo, Dennis, Atmosoedjono, Oemijati & Cross, 1977 cause lymphatic diseases in humans in the tropics, while B. pahangi (Buckley & Edeson, 1956) infects carnivores and causes zoonotic diseases in humans in Malaysia. Wuchereria bancrofti, W. kalimantani Palmieri, Pulnomo, Dennis & Marwoto, 1980 and six out of ten Brugia spp. have been described from Australia, Southeast Asia, Sri Lanka and India...
April 20, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28426724/cloning-retinoid-and-peroxisome-proliferator-activated-nuclear-receptors-of-the-pacific-oyster-and-in-silico-binding-to-environmental-chemicals
#20
Susanne Vogeler, Tamara S Galloway, Michail Isupov, Tim P Bean
Disruption of nuclear receptors, a transcription factor superfamily regulating gene expression in animals, is one proposed mechanism through which pollution causes effects in aquatic invertebrates. Environmental pollutants have the ability to interfere with the receptor's functions through direct binding and inducing incorrect signals. Limited knowledge of invertebrate endocrinology and molecular regulatory mechanisms, however, impede the understanding of endocrine disruptive effects in many aquatic invertebrate species...
2017: PloS One
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