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Daniel S Jones, Sharon A Kessler
Mildew resistance locus O (MLO) proteins are transmembrane proteins that mediate cell-cell communication in plants. We recently demonstrated the importance of subcellular localization to MLO function during pollen tube reception. NORTIA (NTA), the MLO protein involved in this process, localizes to the Golgi of the synergid cell before interaction with the pollen tube. MLO proteins that can substitute for NTA's function in this pathway all partially localize with the same Golgi marker in the synergid cell. In this study, we report that MLO subcellular localization is cell type-dependent, with different distributions of some MLOs observed when ectopically expressed in the epidermal cells of tobacco and Arabidopsis compared to synergids...
October 17, 2017: Plant Signaling & Behavior
Lilian Cruz, Jenny Andrea Arevalo Romero, Mariana Brandão Prado, Tiago G Santos, Marilene Hohmuth Lopes
Extracellular vesicles (EVs) released by mouse embryonic stem cells (mESCs) are considered a source of bioactive molecules that modulate their microenvironment by acting on intercellular communication. Either intracellular endosomal machinery or their derived EVs have been considered a relevant system of signal circuits processing. Herein, we show that these features are found in mESCs. Ultrastructural analysis revealed structures and organelles of the endosomal system such as coated pits and endocytosis-related vesicles, prominent rough endoplasmic reticulum and Golgi apparatus, and multivesicular bodies (MVBs) containing either few or many intraluminal vesicles (ILVs) that could be released as exosomes to extracellular milieu...
October 14, 2017: Stem Cell Reviews
Kanchan Vaswani, Yong Qin Koh, Fatema B Almughlliq, Hassendrini N Peiris, Murray D Mitchell
Exosomes are nanovesicles that play important roles in intercellular communication as they carry information to target cells. Isolation of high purity exosomes will aid in studying the exosomal cargo and quantity as well as how cell-specific messages are carried. We describe a new method incorporating size exclusion chromatography (SEC) to enrich milk-derived exosomes from extracellular vesicles (EVs). This involved the initial isolation of EVs from bovine milk via milk processing and ultracentrifugation; followed by a new method to enrich exosomes using SEC...
October 10, 2017: Reproductive Biology
Guang Hua Li, Shinya Akatsuka, Shan Hwu Chew, Li Jiang, Takahiro Nishiyama, Akihiko Sakamoto, Takashi Takahashi, Mitsuru Futakuchi, Hiromu Suzuki, Kunihiko Sakumi, Yusaku Nakabeppu, Shinya Toyokuni
Oxidative stress including iron excess has been associated with carcinogenesis. The level of 8-oxoguanine, a major oxidatively modified base in DNA, is maintained very low by three distinct enzymes, encoded by OGG1, MUTYH and MTH1. Germline biallelic inactivation of MUTYH represents a familial cancer syndrome called MUTYH-associated polyposis. Here, we used Mutyh-deficient mice to evaluate renal carcinogenesis induced by ferric nitrilotriacetate (Fe-NTA). Although the C57BL/6 background is cancer-resistant, a repeated intraperitoneal administration of Fe-NTA induced a high incidence of renal cell carcinoma (RCC; 26...
October 13, 2017: Pathology International
Mehdi Imani, Hossein Zarei Jaliani, Mohammad Hassan Kheirandish, Mahnaz Azadpour
OBJECTIVES: A newly-introduced protein toxin from a sea anemone, namely fragaceatoxin C is a protein with molecular weight of 20 kDa and pore-forming capability against cell membranes has recently grasped great attentions for its function. In this study, its coding sequence cloned as a fusion protein with His-tag for simple production and rapid purification. MATERIALS AND METHODS: After PCR amplification using NcoI and HindIII-harboring primers, the gene fragment was cloned into pET-28a(+)...
April 2017: Iranian Journal of Basic Medical Sciences
Yuri N Shkryl, Galina N Veremeichik, Dmitriy G Kamenev, Tatiana Y Gorpenchenko, Yulia A Yugay, Dmitriy V Mashtalyar, Aleksander V Nepomnyaschiy, Tatiana V Avramenko, Aleksandr A Karabtsov, Vladimir V Ivanov, Victor P Bulgakov, Sergey V Gnedenkov, Yury N Kulchin, Yury N Zhuravlev
In the present investigation, transgenic tobacco callus cultures and plants overexpressing the silicatein gene LoSilA1 from marine sponge Latrunculia oparinae were obtained and their bioreduction behaviour for the synthesis of silver nanoparticles (AgNPs) was studied. Synthesized nanoparticles were characterized using UV-visible spectroscopy, Fourier transformed infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), atomic flame electron microscopy (AFM) and nanoparticle tracking analysis (NTA)...
October 12, 2017: Artificial Cells, Nanomedicine, and Biotechnology
Haihua Yuan, Rong Deng, Xian Zhao, Ran Chen, Guofang Hou, Hailong Zhang, Yanli Wang, Ming Xu, Bin Jiang, Jianxiu Yu
BACKGROUND: MicroRNAs (miRNAs) are important regulators involved in diverse physiological and pathological processes including cancer. SUMO (small ubiquitin-like modifier) is a reversible protein modifier. We recently found that SUMOylation of TARBP2 and DGCR8 is involved in the regulation of the miRNA pathway. KHSRP is a single stranded nucleic acid binding protein with roles in transcription and mRNA decay, and it is also a component of the Drosha-DGCR8 complex promoting the miRNA biogenesis...
October 11, 2017: Molecular Cancer
Sara Callejón, Ramón Sendra, Sergi Ferrer, Isabel Pardo
Biogenic amines degradation by bacterial laccases is little known, so we have cloned and heterologously expressed, in E. coli, a new laccase from Pediococcus acidilactici CECT 5930 (Lpa5930), a lactic acid bacterium commonly found in foods able to degrade tyramine. The recombinant enzyme has been characterized by physical and biochemical assays. Here we report the optimization of expression and purification procedures of this laccase. DNA encoding sequence of laccase from P. acidilactici was amplified by PCR and cloned into the expression plasmid pET28a for induction by isopropyl-β-D-thiogalactoipyranoside...
2017: PloS One
Jian Wang, Meng-Die Qi, Juan Guo, Ye Shen, Hui-Xin Lin, Lu-Qi Huang
Andrographis paniculata is widely used as medicinal herb in China for a long time and andrographolide is its main medicinal constituent. To investigate the underlying andrographolide biosynthesis mechanisms, RNA-seq for A. paniculata leaves with MeJA treatment was performed. In A. paniculata transcriptomic data, the expression pattern of one member of NAC transcription factor family (ApNAC1) matched with andrographolide accumulation. The coding sequence of ApNAC1 was cloned by RT-PCR, and GenBank accession number was KY196416...
March 2017: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
Beatriz Nogueira Messias Miranda, Wesley Luzetti Fotoran, Fernanda Canduri, Dulce Helena Ferreira de Souza, Gerhard Wunderlich, Emanuel Carrilho
The role of Alpha folate receptors (FRα) in folate metabolism and cancer development has been extensively studied. The reason for this is not only associated to its direct relation to disease development but also to its potential use as a highly sensitive and specific biomarker for cancers therapies. Over the recent years, the crystal structures of human FRα complexed with different ligands were described relying on an expensive and time-consuming production process. Here, we constructed an efficient system for the expression and purification of a human FRα in E...
October 4, 2017: Protein Expression and Purification
A Kariminik, M Saeidi, P Parsia
Glutamate decarboxylase (GAD) converts L-glutamate to g-aminobutyric acid (GABA), which is a non-protein amino acid present in all organisms with some activities including improvement of neurve and cytoskeltal functions. Therefore, GAD is considered as a key molecule to use in molecular therapy of genetical human diseases. Accordingly, cloning of GADs from various plants is an important aim of researchers. The aim of this study was to clone rice (Oryza sativa L.) GADs in Escherichia coli (E.coli) MC 1061 bacterium...
September 30, 2017: Cellular and Molecular Biology
Mikako Tsuji, Arif Md Rashedul Kabir, Masaki Ito, Daisuke Inoue, Kenta Kokado, Kazuki Sada, Akira Kakugo
The water-in-oil emulsion systems have recently been attracting much attention in various fields. However, functionalization of the water-in-oil emulsion systems, which is required for expanding their applications in industry and research, has been challenging. We now demonstrate functionalization of a water-in-oil emulsion system by anchoring a target protein molecule. A microtubule associated motor protein kinesin-1 was successfully anchored to the inner surface of water-in-oil emulsion droplets by employing the specific interaction of the nickel-nitrilotriacetic acid (NTA)-histidine tag...
October 3, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
Jian Zhang, Aiyou Sun, Yuguo Dong, Dongzhi Wei
Prostate apoptosis response-4 (Par-4), an anticancer protein that interacts with cell surface receptor GRP78, can selectively suppress proliferation and induce apoptosis of cancer cells. The core domain of Par-4 (aa 137-195), designated as SAC, is sufficient to inhibit tumor growth and metastasis without harming normal tissues and organs. Nevertheless, the anticancer effects of SAC have not been determined in ovarian cancer cells. Here, we developed a novel method for producing native SAC in Escherichia coli using a small ubiquitin-related modifier (SUMO) fusion system...
October 2, 2017: Applied Biochemistry and Biotechnology
Niloofar Zandvakili, Mohammadreza Zamani, Mostafa Motallebi, Zahra Moghaddassi Jahromi
BACKGROUND: Plants have various defense mechanisms such as production of antimicrobial peptides, particularly pathogenesis related proteins (PR proteins). PR10 family is an essential member of this group, with antifungal, antibacterial and antiviral activities. OBJECTIVE: The goal of this study is to assess the antifungal activity of maize PR10 against some of fungal phytopathogens. MATERIALS AND METHODS: Zea mays PR10 gene (TN-05-147) was cloned from genomic DNA and cDNA and overexpressed in Escherichia coli...
March 2017: Iranian journal of biotechnology
Jun Min, Weiwei Chen, Jinpei Wang, Xiaoke Hu
Compound 2-chloro-5-nitrophenol (2C5NP) is a typical chlorinated nitroaromatic pollutant. To date, the bacteria with the ability to degrade 2C5NP are rare, and the molecular mechanism of 2C5NP degradation remains unknown. In this study, Cupriavidus sp. strain CNP-8 utilizing 2-chloro-5-nitrophenol (2C5NP) and meta-nitrophenol (MNP) via partial reductive pathways was isolated from pesticide-contaminated soil. Biodegradation kinetic analysis indicated that 2C5NP degradation by this strain was concentration dependent, with a maximum specific degradation rate of 21...
2017: Frontiers in Microbiology
Hitesh S Purohit, Lynne S Taylor
PURPOSE: The aim of this research was to study the interplay of solid and solution state phase transformations during the dissolution of ritonavir (RTV) amorphous solid dispersions (ASDs). METHODS: RTV ASDs with polyvinylpyrrolidone (PVP), polyvinylpyrrolidone vinyl acetate (PVPVA) and hydroxypropyl methylcellulose acetate succinate (HPMCAS) were prepared at 10-50% drug loading by solvent evaporation. The miscibility of RTV ASDs was studied before and after exposure to 97% relative humidity (RH)...
September 27, 2017: Pharmaceutical Research
Suresh Susan Aishwarya, Sellamuthu Iyappan, Kamepali Vijaya Lakshmi, Kandathil Narayanan Rajnish
l-Asparaginase is employed in leukaemic treatment and in processing starchy foods. The in silico analysis of Lactobacillus reuteri DSM 20016 reveals the presence of an l-asparaginase gene with theoretical pI value of 4.99. 3D structure prediction was carried out and one model was selected based on the validation scores of 86.293 for ERRAT, 92.10% for VERIFY 3D and Ramachandran plot. Multiple sequence alignment of the protein sequences of l-asparaginases I and II of Escherichia coli, Erwinia chrysanthemum and Homo sapiens shows their sequence similarity...
October 2017: 3 Biotech
Benwei Zhu, Fang Ni, Yun Sun, Zhong Yao
Alginate lyases play an essential role in the production of oligosaccharides by degrading alginate polysaccharide. Although many alginate lyases from various microorganisms have been characterized, reports on alginate lyases with special characteristics and commercial potential are still rather rare. In this study, a new alginate lyase, FsAlgA, was cloned from the deep-sea marine bacterium Flammeovirga sp. NJ-04. The recombinant enzyme was purified on Ni-NTA sepharose and then characterized in detail. It exhibited the highest activity (3343...
September 23, 2017: Extremophiles: Life Under Extreme Conditions
Kridsada Unban, Apinun Kanpiengjai, Saisamorn Lumyong, Thu-Ha Nguyen, Dietmar Haltrich, Chartchai Khanongnuch
Gene encoding cyclomaltodextrinase (Cdx) from amylolytic lactic acid bacterium Enterococcus faecium K-1 was cloned and nucleotide sequence was analyzed. The open-reading frame consisted of 1767bp encoding 588 deduced amino acids. Consequently, four typically conserved regions of the glycoside hydrolase family 13 were revealed; however, nine exceeding amino acids (DSYQMTDVP) were found at the 282-290 position in comparison to previously reported cyclomaltodextrinases. This difference is believed to have an influence on the substrate specificity of this enzyme...
September 18, 2017: International Journal of Biological Macromolecules
Yong Qin Koh, Fatema B Almughlliq, Kanchan Vaswani, Hassendrini N Peiris, Murray D Mitchell
Exosomes are a subset of extracellular vesicles (EVs) that have important roles in intercellular communication. They contain and carry bioactive molecules within their membranes which are delivered to target cells. Reproducible isolation and enrichment of these exosomes will aid in evaluation of cellular communication. We present an approach that involved the pre-processing of plasma, combined with ultracentrifugation (UC) and size exclusion chromatography (SEC) to isolate EVs and subsequently enrich exosomes...
January 1, 2018: Frontiers in Bioscience (Landmark Edition)
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