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https://www.readbyqxmd.com/read/28936836/-molecular-cloning-and-characterization-of-two-kind-of-heat-shock-protein-gene-from-polyporus-umbellatus
#1
Meng-Meng Liu, Yong-Mei Xing, Shun-Xing Guo
With RT-PCR approaches, the full-length cDNA of two heat shock protein genes were cloned from total RNA of the Polyporus umbellatus sclerotium. The full open reading frame cDNA sequence of the Hsp90 was 2 091 bp, encoding 696 amino acid residues with a predicted molecular mass of 78.9 kDa. The full open reading frame cDNA sequence of the Hsp70 was 1 944 bp, encoding 647 amino acid residues with a predicted molecular mass of 70.5 kDa. The Hsp90 and Hsp70 protein contained the conservative structure domain, respectively...
December 2016: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28935539/molecular-structure-of-cyclomaltodextrinase-derived-from-amylolytic-lactic-acid-bacterium-enterococcus-faecium-k-1-and-properties-of-recombinant-enzymes-expressed-in-escherichia-coli-and-lactobacillus-plantarum
#2
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
https://www.readbyqxmd.com/read/28935259/module-based-systematic-construction-of-plasmids-for-episomal-gene-expression-in-fission-yeast
#3
Keita Kiriya, Hayato Tsuyuzaki, Masamitsu Sato
The fission yeast Schizosaccharomyces pombe is a powerful model organism for cell biology and molecular biology, as genetic manipulation is easily achieved. Introduction of exogenous genes cloned in episomal plasmids into yeast cells can be done through well-established transformation methods. For expression of genes in S. pombe cells, the multi-copy plasmid pREP1 and its derivatives, including pREP41 and pREP81, have been widely used as vectors. Although recent advancement of technology brought a number of useful genetic elements such as new promoters, selection marker genes and fluorescent protein tags, introduction of those elements into conventional pREP1 requires a large commitment of both time and effort because cloning procedures need to be repeated until the final products are constructed...
September 18, 2017: Gene
https://www.readbyqxmd.com/read/28933241/characterization-of-human-s-adenosyl-homocysteine-hydrolase-in-vitro-and-identification-of-its-potential-inhibitors
#4
Weiwei Hao, Yanhua Li, Qiuli Shan, Tian Han, Wencheng Li, Sheng He, Kongkai Zhu, Yumei Li, Xiaojun Tan, Jinsong Gu
Human S-adenosyl-homocysteine hydrolase (SAHH, E.C.3.3.1.1) has been considered to be an attractive target for the design of medicines to treat human disease, because of its important role in regulating biological methylation reactions to catalyse the reversible hydrolysis of S-adenosylhomocysteine (SAH) to adenosine (Ado) and l-homocysteine (Hcy). In this study, SAHH protein was successfully cloned and purified with optimized, Pichia pastoris (P. pastoris) expression system. The biological activity results revealed that, among the tested compounds screened by ChemMapper and SciFinder Scholar, 4-(3-hydroxyprop-1-en-1-yl)-2-methoxyphenol (coniferyl alcohol, CAS: 458-35-5, ZINC: 12359045) exhibited the highest inhibition against rSAHH (IC50= 34 nM)...
December 2017: Journal of Enzyme Inhibition and Medicinal Chemistry
https://www.readbyqxmd.com/read/28932628/a-novel-in-silico-framework-to-improve-mhc-i-epitopes-and-break-the-tolerance-to-melanoma
#5
Cristian Capasso, Aniket Magarkar, Victor Cervera-Carascon, Manlio Fusciello, Sara Feola, Martin Muller, Mariangela Garofalo, Lukasz Kuryk, Siri Tähtinen, Lucio Pastore, Alex Bunker, Vincenzo Cerullo
Tolerance toward tumor antigens, which are shared by normal tissues, have often limited the efficacy of cancer vaccines. However, wild type epitopes can be tweaked to activate cross-reactive T-cell clones, resulting in antitumor activity. The design of these analogs (i.e., heteroclitic peptides) can be difficult and time-consuming since no automated in silico tools are available. Hereby we describe the development of an in silico framework to improve the selection of heteroclitic peptides. The Epitope Discovery and Improvement System (EDIS) was first validated by studying the model antigen SIINFEKL...
2017: Oncoimmunology
https://www.readbyqxmd.com/read/28932329/molecular-characterization-of-klebsiella-pneumoniae-clinical-isolates-with-elevated-resistance-to-carbapenems
#6
Rasha Barwa, Mona Shaaban
BACKGROUND: Emergence of carbapenems-resistant K. pneumoniae represents a serious challenge for antimicrobial therapy. OBJECTIVE: The aim of this research is to determine different mechanisms mediating the emergence of K. pneumoniae isolates with high-level carbapenem resistance. METHOD: A total of 80 K. pneumoniae isolates were purified from sputum and urine specimens. The minimum inhibitory concentrations (MICs) of imipenem and meropenem were determined by broth microdilution method...
2017: Open Microbiology Journal
https://www.readbyqxmd.com/read/28931519/beta-cell-replacement-in-mice-using-human-type-1-diabetes-nuclear-transfer-embryonic-stem-cells
#7
Lina Sui, Nichole Danzl, Sean R Campbell, Ryan Viola, Damian Williams, Yuan Xing, Yong Wang, Neil Phillips, Greg Poffenberger, Bjarki Johannesson, Jose Oberholzer, Alvin C Powers, Rudolph L Leibel, Xiaojuan Chen, Megan Sykes, Dieter Egli
Beta cells derived from stem cells hold great promise for cell replacement therapy for diabetes. Here we examine the ability of nuclear transfer embryonic stem cells (NT-ES) derived from a type 1 diabetes patient to differentiate into beta cells, and provide a source of autologous islets for cell replacement. NT-ES cells differentiate in vitro with an average efficiency of 55% into C-peptide-positive cells, expressing markers of mature beta cells, including MAFA and NKX6.1. Upon transplantation in immunodeficient mice, grafted cells form vascularized islet-like structures containing MAFA/C-peptide-positive cells...
September 20, 2017: Diabetes
https://www.readbyqxmd.com/read/28931412/molecular-characterization-of-the-duck-enteritis-virus-us10-protein
#8
Daixi Zhang, Maoyin Lai, Anchun Cheng, Mingshu Wang, Ying Wu, Qiao Yang, Mafeng Liu, Dekang Zhu, Renyong Jia, Shun Chen, Kunfeng Sun, Xinxin Zhao, Xiaoyue Chen
BACKGROUND: There is little information regarding the duck enteritis virus (DEV) US10 gene and its molecular characterization. METHODS: Duck enteritis virus US10 was amplified and cloned into the recombinant vector pET32a(+). The recombinant US10 protein was expressed in Escherichia coli BL21 cells and used to immunize rabbits for the preparation of polyclonal antibodies. The harvested rabbit antiserum against DEV US10 was detected and analyzed by agar immunodiffusion...
September 20, 2017: Virology Journal
https://www.readbyqxmd.com/read/28930002/investigation-of-outbreaks-of-salmonella-enterica-serovar-typhimurium-and-its-monophasic-variants-using-whole-genome-sequencing-denmark
#9
Pernille Gymoese, Gitte Sørensen, Eva Litrup, John Elmerdal Olsen, Eva Møller Nielsen, Mia Torpdahl
Whole-genome sequencing is rapidly replacing current molecular typing methods for surveillance purposes. Our study evaluates core-genome single-nucleotide polymorphism analysis for outbreak detection and linking of sources of Salmonella enterica serovar Typhimurium and its monophasic variants during a 7-month surveillance period in Denmark. We reanalyzed and defined 8 previously characterized outbreaks from the phylogenetic relatedness of the isolates, epidemiologic data, and food traceback investigations. All outbreaks were identified, and we were able to exclude unrelated and include additional related human cases...
October 2017: Emerging Infectious Diseases
https://www.readbyqxmd.com/read/28929648/-cloning-and-expression-analysis-of-tocopherol-cyclase-gene-in-carthamus-tinctorius
#10
Li-Li Guan, Qi Cui, Yu-Tong Han, Yun-Yun Wu, Ren-Ge Hu, Tian-Yao Gu, Hai-Yan Li, Xiao-Kun Li
The tocopherol cyclase was one of the key enzymes in plant vitamin E biosynthesis pathway. According to the study of Carthamus tinctorius transcriptome data,the Tocopherol cyclase gene was obtained using RT-PCR techniques and named CtTC . Bioinformatics analysis showed theopen reading frame (ORF)of CtTC was 1 524 bp. The putative protein contained 507 amino acids with a predicted molecular mass of 62.9 kDa and theoretically isoelectric point was 5.01.Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide...
October 2016: Zhongguo Zhong Yao za Zhi, Zhongguo Zhongyao Zazhi, China Journal of Chinese Materia Medica
https://www.readbyqxmd.com/read/28929019/heterologous-expression-of-plasmodium-vivax-apical-membrane-antigen-1-pvama1-for-binding-peptide-selection
#11
Ching Hoong Chew, Yvonne Ai Lian Lim, Kek Heng Chua
BACKGROUND: Plasmodium is an obligate intracellular parasite. Apical membrane antigen 1 (AMA1) is the most prominent and well characterized malarial surface antigen that is essential for parasite-host cell invasion, i.e., for sporozoite to invade and replicate within hepatocytes in the liver stage and merozoite to penetrate and replicate within erythrocytes in the blood stage. AMA1 has long served as a potent antimalarial drug target and is a pivotal vaccine candidate. A good understanding of the structure and molecular function of this Plasmodium protein, particularly its involvement in host-cell adhesion and invasion, is of great interest and hence it offers an attractive target for the development of novel therapeutics...
2017: PeerJ
https://www.readbyqxmd.com/read/28928395/stochastic-priming-and-spatial-cues-orchestrate-heterogeneous-clonal-contribution-to-mouse-pancreas-organogenesis
#12
Hjalte List Larsen, Laura Martín-Coll, Alexander Valentin Nielsen, Christopher V E Wright, Ala Trusina, Yung Hae Kim, Anne Grapin-Botton
Spatiotemporal balancing of cellular proliferation and differentiation is crucial for postnatal tissue homoeostasis and organogenesis. During embryonic development, pancreatic progenitors simultaneously proliferate and differentiate into the endocrine, ductal and acinar lineages. Using in vivo clonal analysis in the founder population of the pancreas here we reveal highly heterogeneous contribution of single progenitors to organ formation. While some progenitors are bona fide multipotent and contribute progeny to all major pancreatic cell lineages, we also identify numerous unipotent endocrine and ducto-endocrine bipotent clones...
September 19, 2017: Nature Communications
https://www.readbyqxmd.com/read/28928248/carbonic-anhydrase-iii-protects-osteocytes-from-oxidative-stress
#13
Chao Shi, Yuhei Uda, Christopher Dedic, Ehab Azab, Ningyuan Sun, Amira I Hussein, Christopher A Petty, Keertik Fulzele, Maria C Mitterberger-Vogt, Werner Zwerschke, Renata Pereira, Kunzheng Wang, Paola Divieti Pajevic
Osteocytes are master orchestrators of bone remodeling; they control osteoblast and osteoclast activities both directly via cell-to-cell communication and indirectly via secreted factors, and they are the main postnatal source of sclerostin and RANKL (receptor activator of NF-kB ligand), two regulators of osteoblast and osteoclast function. Despite progress in understanding osteocyte biology and function, much remains to be elucidated. Recently developed osteocytic cell lines-together with new genome editing tools-has allowed a closer look at the biology and molecular makeup of these cells...
September 19, 2017: FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology
https://www.readbyqxmd.com/read/28928065/molecular-cloning-and-biophysical-characterization-of-cxcl3-chemokine
#14
Khushboo Gulati, Krishnakant Gangele, Nipanshu Agarwal, Minal Jamsandekar, Dinesh Kumar, Krishna Mohan Poluri
CXCL3 is a neutrophil activating chemokine that belongs to GRO subfamily of CXC chemokines. GRO chemokine family comprises of three chemokines GRO α (CXCL1), GROβ (CXCL2), and GRO γ (CXCL3) that arose as a result of gene duplication events during the course of chemokine evolution. Studies reported that the primary sequences of GRO chemokines are highly similar, but performs several protein specific functions in addition to their common property of neutrophil trafficking. However, the molecular basis for their differential functions has not well understood...
September 16, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28926428/dna-methyltransferase-inhibitors-in-myeloid-cancer-clonal-eradication-or-clonal-differentiation
#15
Andreas Due Ørskov, Kirsten Grønbæk
DNA methyltransferase inhibitors, so-called hypomethylating agents (HMAs), are the only drugs approved for the treatment of higher-risk myelodysplastic syndromes and are widely used in this context. However, it is still unclear why some patients respond to HMAs, whereas others do not. Recent sequencing efforts have identified molecular disease entities that may be specifically sensitive to these drugs, and many attempts are being made to clarify how HMAs affect the malignant clone during treatment. Here, we review the most recent data on the clinical effects of HMAs in myeloid malignancies...
September 2017: Cancer Journal
https://www.readbyqxmd.com/read/28924554/molecular-cloning-and-functional-identification-of-sterol-c24-methyltransferase-gene-from-tripterygium-wilfordii
#16
Hongyu Guan, Yujun Zhao, Ping Su, Yuru Tong, Yujia Liu, Tianyuan Hu, Yifeng Zhang, Xianan Zhang, Jia Li, Xiaoyi Wu, Luqi Huang, Wei Gao
Sterol C24-methyltransferase (SMT) plays multiple important roles in plant growth and development. SMT1, which belongs to the family of transferases and transforms cycloartenol into 24-methylene cycloartenol, is involved in the biosynthesis of 24-methyl sterols. Here, we report the cloning and characterization of a cDNA encoding a sterol C24-methyltransferase from Tripterygium wilfordii (TwSMT1). TwSMT1 (GenBank access number KU885950) is a 1530 bp cDNA with a 1041 bp open reading frame predicted to encode a 346-amino acid, 38...
September 2017: Acta Pharmaceutica Sinica. B
https://www.readbyqxmd.com/read/28924501/identification-and-analysis-of-cyp450-genes-from-transcriptome-of-lonicera-japonica-and-expression-analysis-of-chlorogenic-acid-biosynthesis-related-cyp450s
#17
Xiwu Qi, Xu Yu, Daohua Xu, Hailing Fang, Ke Dong, Weilin Li, Chengyuan Liang
BACKGROUND: Lonicera japonica is an important medicinal plant that has been widely used in traditional Chinese medicine for thousands of years. The pharmacological activities of L. japonica are mainly due to its rich natural active ingredients, most of which are secondary metabolites. CYP450s are a large, complex, and widespread superfamily of proteins that participate in many endogenous and exogenous metabolic reactions, especially secondary metabolism. Here, we identified CYP450s in L...
2017: PeerJ
https://www.readbyqxmd.com/read/28924030/altered-regulation-of-the-diguanylate-cyclase-yaic-reduces-production-of-type-1-fimbriae-in-a-pst-mutant-of-uropathogenic-e-coli-cft073
#18
Sébastien Crépin, Gaëlle Porcheron, Sébastien Houle, Josée Harel, Charles M Dozois
The pst gene cluster encodes the phosphate specific transport system (Pst). Inactivation of the Pst system constitutively activates the two-component regulatory system PhoBR and attenuates virulence of pathogenic bacteria. In uropathogenic E. coli strain CFT073, attenuation by inactivation of pst is predominantly attributed to the decreased expression of type 1 fimbriae. However, the molecular mechanisms connecting the Pst system and type 1 fimbriae are unknown. To address this, a transposon library was constructed in the pst mutant, and clones were tested for a regain in type 1 fimbriae production...
September 18, 2017: Journal of Bacteriology
https://www.readbyqxmd.com/read/28920412/molecular-cloning-purification-expression-and-characterization-of-%C3%AE-1-4-endoglucanase-gene-cel5a-from-eubacterium-cellulosolvens-sp-isolated-from-holstein-steer-s-rumen
#19
Tansol Park, Seongwon Seo, Teaksoon Shin, Byung-Wook Cho, Seongkeun Cho, Byeongwoo Kim, Jong K Ha, Jakyeom Seo
Objective: This study was conducted to isolate the cellulolytic microorganism from a rumen of Holstein steer and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods: To isolate anaerobic microbes having endoglucanase, the rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 99% similarity with Eubacterium cellulosolvens Ce2 (Accession number: AY178842.1). The Cel5A from isolated E. cellulosolvens sp...
September 18, 2017: Asian-Australasian Journal of Animal Sciences
https://www.readbyqxmd.com/read/28919972/cytogenetic-maps-of-homoeologous-chromosomes-a-h01-and-d-h01-and-their-integration-with-the-genome-assembly-in-gossypium-hirsutum
#20
Yuling Liu, Zhen Liu, Renhai Peng, Yuhong Wang, Zhongli Zhou, Xiaoyan Cai, Xingxing Wang, Zhenmei Zhang, Kunbo Wang, Fang Liu
Cytogenetic maps of Gossypium hirsutum (Linnaeus, 1753) homoeologous chromosomes Ah01 and Dh01 were constructed by fluorescence in situ hybridization (FISH), using eleven homoeologous-chromosomes-shared bacterial artificial chromosomes (BACs) clones and one chromosome-specific BAC clone respectively. We compared the cytogenetic maps with the genetic linkage and draft genome assembly maps based on a standardized map unit, relative map position (RMP), which allowed a global view of the relationship of genetic and physical distances along each chromosome, and assembly quality of the draft genome assembly map...
2017: Comparative Cytogenetics
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