gene identification cellular localization prediction and functional characterization for vaccine candidate selection
Tài liệu Báo cáo khoa học: Molecular and functional characterization of adenylate kinase 2 gene from Leishmania donovani pdf
... Asian and Mediterranean countries [15] This paper describes the cloning and functional characterization of an AK gene from a genomic library of L donovani as well as its expression and molecular characterization ... protocol Isolation of a L donovani AK gene fragment by PCR To generate a DNA probe for the isolation of the L donovani AK gene, PCR was carried out using degenerate oligonucleotides based on conserved ... (Amersham) and prehybridized for h at 42 °C in · Denhardt’s solution, · NaCl/Cit, 50 mM sodium phosphate (pH 6.5), 100 lgÆmL)1 salmon sperm DNA, and 50% formamide Membranes were hybridized for 16–20...
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Báo cáo khoa học: "Structural and functional characterization of the 5’ upstream region of a glutamine synthetase gene from Scots pine" pdf
... introns is usually between 70% described for dicot plants and around 60% for monocot plants [22] Unfortunately not many data are available for gymnosperm genes, however the introns in the GS1 clone ... subcloned into the plasmid pGEM-3Z to generate the clone pGS217 and used for analysis and sequencing 2.2 Fusions of the 5’region of GS1a to the GUS reporter gene The 981 bp sequence upstream of ... were transformed individually in Agrobacterium tumefaciens LBA4104 2.3 Transient and stable transformation with the gene constructs A Biolistic PDS-1000/He apparatus from Bio-Rad was used for particle...
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Báo cáo y học: "Identification and functional characterization of cis-regulatory elements in the apicomplexan parasite Toxoplasma gond" pptx
... Figure Candidate motifs identified upstream of glycolytic genes, upstream location, site-directed mutagenesis and results of reporter assays Candidate motifs identified upstream of glycolytic genes, ... mutagenized Promoter Candidate motifs identified upstream of the nucleotide biosynthetic genes, upstream location, site-directed mutagenesis and results of reporter assays Figure Candidate motifs ... Promoter M2AP Figure assays Candidate motifs identified upstream of the micronemal protein-encoding genes, upstream location, site-directed mutagenesis and results of reporter Candidate motifs identified...
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Biochemical identification and functional characterization of microrna target interactions in growth control and cancer transformation
... (representative reviews in (Iorio and Croce, 2012; Mendell and Olson, 2012; Rottiers and Naar, 2012)) My PhD project is focused on identification and functional characterizations of miRNA-target ... use and widely applicable for a variety of cell types, genetic manipulation of microRNA and target interaction in vivo is possible The microRNA and/ or wildtype and mutant target sequence transgenes ... Benjamini and Hochberg 2.8 miRNA Target Site Prediction Prediction of potential miRNA binding sites for both S2 expressed and all D melanogaster miRNAs was performed similarly to that described for...
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Cellular and molecular control of skeleton formation in fish insights from osteoblast ablation and functional characterization of lrp5 and SOst
... Singapore and the Department of Biological Sciences for my admission into the graduate programme and the generous scholarship Thanks also for creating an excellent environment and for providing ... as well as for his wonderful mentoring and guidance His helpful advice and our fruitful discussions helped me a lot to accomplish my candidature I wish him and his family all the best for the future ... Strand cDNA Synthesis Kit (Fermentas) RNA was diluted with H2O to a volume of 11µl and 1µl of random hexamer primer was added For annealing the solution was incubated at 70°C for minutes and...
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Genomic organization and functional characterization of a novel cancer associated gene u0 44
... as a candidate tumor suppressor gene for the human brain, lung and digestive tract cancers; it differs substantially from conventional tumor suppressors and is in fact potentially multifunctional ... DISCUSSION 5.1 Human UO-44 gene 5.1.1 Features in HuUO-44 Gene 5.1.2 The Origin of Human HuUO-44 Isoforms 5.1.3 UO-44, a Multifunctional Protein 5.2 Rat Model for Further Characterization of UO-44 ... proliferation, chemotaxis and transformation It has been known to play a key role in cell-cell communication for normal development and also during pathogenesis (Rosenkranz and Kazlauskas, 1999;...
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Group 2 allergens from dust mite epitope mapping and functional characterization of der p 2, and identification of a paralogue of der f 2
... treatment for allergic diseases 12 1.2 Aims 14 Chapter 2: Materials and methods 16 2.1 Cloning, mutagenesis, DNA sequencing and gene characterization 16 2.1.1 Sub-cloning and site-directed mutagenesis ... were then evaluated for their efficacy as hypoallergen vaccine candidates Mutants E102A and K96A (which resulted in an unfolded protein) were potential hypoallergen vaccine candidates as they demonstrated ... profile 15 Chapter 2: Materials and methods 2.1 Cloning, mutagenesis, DNA sequencing and gene characterization 2.1.1 Sub-cloning and site-directed mutagenesis DNA encoding for allergens were amplified...
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Tài liệu Báo cáo khoa học: Molecular modeling and functional characterization of the monomeric primase–polymerase domain from the Sulfolobus solfataricus plasmid pIT3 doc
... 60, 70 and 80 °C between 190 and 260 nm with a step increase of 0.2 nm, and a bandwidth of nm Thermal stability of Rep245 was measured by incubating the protein at 60, 70 and 80 °C for 30 and then ... follows: Na-acetate (pH 5.0, 5.4 and 5.8), Tris ⁄ HCl (pH 6.5, 7.0, 7.5 and 8.0) and glycine ⁄ NaOH (pH 8.6, 9.0 and 9.6) Reactions were carried out for 30 at 65 °C and the products resolved by denaturing ... to serve as ligands for divalent cations and as tags revealing the presence of DNA polymerases in the active site [18–20] In this study, we report on the structural and functional characterization...
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Báo cáo khoa học: Molecular and functional characterization of novel CRFR1 isoforms from the skin pptx
... (Table 1) For example, band (48 kDa) corresponds to the expected 47.4 kDa for pCRFR1f-V5; band (43 kDa) to 43.5 kDa molecular mass for pCRFR1g-V5; band 11 (16 kDa) to 15.1 kDa molecular mass for CRHR1e1 ... cells cotransformed by constructs containing CRE, CaRe, AP1, NF-jB and SRE elements and different CRFR1 isoforms (A) Stimulation by CRF (B) Stimulation by CRF, forskolin and PMA (TPA) and inhibition ... pepstatin and mM amino-ethyl benzene sulfonyl fluoride (AEBSF) Cellular homogenates were centrifuged at 16 000 g for 10 at °C, and the supernatants were removed and stored at )80 °C for further...
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Báo cáo khoa học: Molecular and functional characterization of a novel splice variant of ANKHD1 that lacks the KH domain and its role in cell survival and apoptosis docx
... Swiss Institute of Bioinformatics were used for in sillico characterization of VBARP, including prediction of its subcellular localization (psort: http://www.psort.org), prediction of post-translational ... 2B that VBARP-L and VBARP-S are coded by these specific exons VBARP-L and VBARP-S code for precursor proteins of 627 and 435 amino acids and with calculated peptide masses of 69 and 49 kDa, respectively ... modifications and ⁄ or splice variants Subcellular distribution of VBARP To determine the subcellular localization of VBARP isoforms, His-tagged VBARP plasmids were transfected into HeLa cells, and the...
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Báo cáo khoa học: Sulfation of hydroxychlorobiphenyls Molecular cloning, expression, and functional characterization of zebrafish SULT1 sulfotransferases docx
... compounds and 50 mM Mops at pH 7.0 were used To evaluate their thermostability, the zebrafish STs were first incubated for 15 at, respectively, 20, 28, 37, 43 and 48 °C, and then assayed for their ... buffer and heated for at 100 °C, were separated by SDS/PAGE and electrotransferred onto an Immobilon-P membrane [18] The blotted membrane was blocked with 5% nonfat dried milk in NaCl/Pi for h and ... zebrafish SULT1 ST1 and ST2 mRNAs and (B) Western blot analysis of zebrafish SULT1 ST1 protein (A) Detection of zebrafish SULT1 ST1 and ST2 mRNAs in cultured zebrafish cells (lanes and 3) and whole zebrafish...
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Báo cáo khoa học: Identification and functional characterization of a novel barnacle cement protein pptx
... polypeptides were predicted to be 16 995.52 Da (Table 1) and 5.8 for Mrcp-19k, 17 336.27 Da and 10.3 for Balcp-19k, and 16 841.99 Da and 10.3 for Bicp-19k, respectively The molecular mass of Mrcp-19k ... biotechnology and material science, e.g development of adsorbents for various material surfaces, of support for protein alignment on a solid surface [25–27], and of underwater adhesives for surgical ... improvisus and B albicostatus were collected from Miyako Bay (Iwate), Yodo River (Osaka) and Shimizu Bay (Shizuoka, Japan), respectively RNA and DNA manipulation was generally performed as described...
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Báo cáo khoa học: Structural and functional characterization of human Iba proteins ppt
... EF-hand 2, although EF-hand does not bind Ca2+ Therefore, it may be questioned if the rearrangement and dimerization of the Iba proteins are indeed caused by Ca2+ binding Cellular localization and ... Dinh-Trung Pham and Janett Tischer for technical assistance and Ulrich Harttig for help in target selection This study was funded by the German Federal Ministry for Education and Research (BMBF) ... binding was observed for Iba1 and Iba2 Cellular localization and recruitment to sites of Shigella invasion We expressed cyan uorescent protein (CFP)-tagged Iba2 in HeLa cells and found that the...
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Báo cáo Y học: Structural and functional characterization of a C-type lectin-like antifreeze protein from rainbow smelt (Osmerus mordax) potx
... Locke for trypsin, and Neil Ross for time on his fluorimeter and patient instruction in software use for the instrument We are grateful to A/F Protein Canada Inc for their generous donation of ... colleagues for their kind assistance, including Robert Richards for helpful discussion, Shawna MacKinnon for use of her HPLC, Denise LeBlanc for loan of a size exclusion HPLC column, Steve Locke for ... both untreated and deglycosylated smelt AFP were digested with Glu-C and trypsin in the presence and absence of Ca2+ Both enzymes generated some proteolysis near the N-terminus and/ or C-terminus...
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Báo cáo khoa học: Purification and functional characterization of human 11b hydroxylase expressed in Escherichia coli doc
... compared to 45 h for the pTrc99A ⁄ JM109 system used for the expression of rat CYP11B1 and CYP11B2 [15] The expressed form of hCYP11B1 was stable in the presence of detergents and glycerol and highly ... isoforms (cortisol and aldosterone formation) are performed by a single protein CYP11B1 deficiency results in decreased cortisolproduction leading subsequently to an elevated plasma ACTH level, and ... absorption band [22] Upon addition of substrate, the signal of the positive CD band at 290 nm and the negative band at 386 nm decreased (Fig 4B) A similar observation was described for substrate...
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Báo cáo khoa học: Cloning and functional characterization of Arabidopsis thaliana D-amino acid aminotransferase – D-aspartate behavior during germination pdf
... determinations were performed for each time point, and essentially similar results were obtained The data shown represent the results obtained in an experiment Gene cloning and functional characterization ... parameters for these activities were determined, and are shown in Table In the transamination conversion of amino acid to a-ketoglutarate, Km and Vmax values for d-Asp and d-Ala were 2.3 and mm, and ... produce d-Ala were mm and 3.3 lmolÆmin)1Æmg)1 protein, respectively (Table 1) Therefore the affinity for d-Ala is higher than that for d-Glu, and Vmax is higher for d-Ala than for d-Glu, indicating...
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Báo cáo khoa học: Identification and functional characterization of an aggregation domain in long myosin light chain kinase ppt
... encoded by a candidate gene for limb-girdle muscular dystrophy Hum Mol Genet 8, 1329–1336 35 Parast MM & Otey CA (2000) Characterization of palladin, a novel protein localized to stress fibers and cell ... tests for transgenic muscles (A, A¢) (diaphragm and EDL muscle): mean tetanic force (± SEM) during repetitive isometric activation , wild-type; d, 4Ig-Tg (B, B¢) (diaphragm and EDL muscle) and ... ice for 30 Following this, they were centrifuged at 8064 g for 10 min, and the pellet (P1) and supernatant (S1) were collected P1 was further dissolved in lysis buffer containing 2% SDS, and...
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Báo cáo khoa học: Expression and functional characterization of P2Y1 and P2Y12 nucleotide receptors in long-term serum-deprived glioma C6 cells ppt
... phosphorylation, and P2Y1 and P2Y12 receptor protein expression and functional activity Examination of the effects of specific pharmacological agents (a single agonist and two specific antagonists of P2Y1 and ... serum for up to 96 h before the experiment For morphology changes, the cells were cultured in MEM with 10% (v ⁄ v) fetal bovine serum, or for 48, 72 and 96 h in MEM without fetal bovine serum, and ... cells starved for 96 h and incubation for the next 24 h (Fig 3, M-) increased the Fig Cell cycle analysis performed on propidium iodide-stained glioma C6 cells Each phase (Go ⁄ Gi, S and G2 ⁄ M)...
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Báo cáo khóa học: Purification and functional characterization of insecticidal sphingomyelinase C produced by Bacillus cereus ppt
... (VS and MA, MS and VA), mM MgSO4 and 0.2 mM dNTP mixture in a 50-lL solution at 94 °C for followed by 30 cycles of 94 °C for 15 s, 50 °C for 30 s, and 68 °C for The second-round PCR was performed ... primers (VS and VA), mM MgSO4 and 0.2 mM dNTP mixture in a 50-lL solution at 94 °C for followed by 35 cycles of 94 °C for 15 s, 50 °C for 30 s, and 68 °C for min, yielding a single band of predicted ... 0.2 mM dNTP and 100 ng genomic DNA by the following protocol (final volume, 50 lL): 94 °C for followed by 30 cycles of 94 °C for 15 s, 50 °C for 30 s, and 68 °C for The amplified DNA band was purified...
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Báo cáo khoa học: Cloning and functional characterization of Phaeodactylum tricornutum front-end desaturases involved in eicosapentaenoic acid biosynthesis doc
... Germany, Stratagene) After two rounds of screening, several clones were isolated and the longest ones sequenced on both strands Functional characterization in S cerevisiae For functional characterization, ... We thank Dr T K Zank for providing the pY2PSE1 clone and BASF Plant Science GmbH (Ludwigshafen, Germany) for providing the k-ZAP Express library and for performing the random sequencing REFERENCES ... frames of PtD6 and PtD5 were cloned in the yeast expression vectors pVT102-U [26] and pESC-LEU (Strategene), respectively, and these constructs and the empty vectors were transformed into the...
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