DNA purification
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In a previous study, a gene GL0694641 coding for endoglucanase containing 3 domains GH5- CBM72-CBM72 was exploited from metagenomic DNA data of bacteria in Vietnamese goats’ rumen. The gene (eg5) encoding the mature enzyme (without signal peptide coding sequence) was optimized codons, artificially synthesized, and inserted into the pET22b(+) vector at NcoI and XhoI to generate expression vector pET22-eg5 for expression of the gene in Eschrichia coli.
11p vicaptainmarvel 21-04-2023 5 2 Download
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We examined the properties of the nanocomposite γ-Fe2O3@Chi@Pani as an adsorbent of deoxyribonucleic acid (DNA). As a model system, we used an aqueous solution of salmon sperm DNA, whose decreasing concentration was followed by monitoring the 260 nm UV–vis absorption.
9p vihermione 06-01-2023 7 2 Download
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Protein purification by affinity chromatography relies primarily on the interaction of a fused-tag to the protein of interest. Here, we describe a tag-free affinity method that employs functional selection interactions to a broad range of proteins.
9p viginny 23-12-2022 12 3 Download
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Multicolumn Countercurrent Solvent Gradient Purification (MCSGP) is a continuous chromatography technique used to maximize purification yields compared to traditional batch purification methods. Here we apply MCSGP for the reversed phase purification of a N-acetylgalactosamine (GalNAc)-cluster-conjugated DNA-LNA gapmer oligonucleotide therapeutic using a twin-column chromatography system.
8p viginny 23-12-2022 12 3 Download
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A high density lipoprotein (HDL)-linked enzyme with antioxidant and antiatherogenic properties, paraoxonase1(PON1), prevents the formation of atherosclerotic lesions in humans. In the present study, a recombinant hPON1 gene was produced using a small ubiquitin-related modifier (SUMO) fusion protein expression system. To that end, the hPON1 gene was amplified from human liver-ready cDNA, cloned into the expression vector pET SUMO, and expressed in Escherichia coli BL21 (DE3).
13p langthannam 29-12-2021 8 1 Download
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In this study, the DNA fragment encoding for the C-terminus domain of the AopB from Aeromonas hydrophila AH-1 was cloned into pET-M expression vector and expressed in Escherichia coli BL21 (DE3) host cells. The recombinant AopB-C-terminus domain was successfully purified using immobilized nickel affinity chromatography as a soluble form. Crosslinking analysis among AopB-C-terminus molecules in solution showed that this domain existed as a mixture of tetramer, trimer, dimer, and monomer forms.
6p guitaracoustic02 08-12-2021 18 3 Download
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The present study describes the simultaneous expression of thermostable industrial alpha (α) and beta (β) amylase enzymes that have been used widely in starch industry. Genomic DNA of Bacillus stearothermophilus DSM 22 strain for α amylase and, Thermoanaerobacterium (Clostridium) thermosulfurogenes DSM 2229 strain for β amylase were used as gene sources. Both genes were ligated into pETDuet-1 expression vector separately and resulting recombinant vectors were transformed into Escherichia coli BL21 competent cells by electroporation.
7p thiencuuchu 27-11-2021 17 1 Download
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Chromatin immunoprecipitation-sequencing (ChIP-seq) is a widely used epigenetic approach for investigating genome-wide protein-DNA interactions in cells and tissues. The approach has been relatively well established but several key steps still require further improvement.
10p vilarryellison 29-10-2021 7 0 Download
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While numerous subpopulations of BM-MSCs have been identified, the relevance of these findings regarding the functional properties remains mostly unclear. With regards to attempts of enhancing differentiation results by preselecting certain MSC subtypes, we have evaluated the efficiency of CD146 purification during expansion, and evaluated whether these measures enhanced MSC differentiation results.
10p viannito2711 20-04-2021 10 2 Download
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Objective of this study was to identify selected species of Cordyceps genus using DNA barcodes. Seven strains of Cordyceps were collected. Total DNA extraction and purification, PCR amplification and DNA sequencing were performed with standard chemicals and kits. The candidate ITS1-5.8S-ITS2 region was amplified and sequenced. Data were analyzed using Bioedit 7.2.6 and MEGA 7 softwares. Analysis of seven obtained DNA barcode sequences of collected samples revealed that the ITS1- 5.8S-ITS2 region provided high species discriminating power for Cordyceps genus.
8p nguyenxuankha_bevandan 14-08-2020 17 1 Download
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Oropharyngeal Squamous Cell Carcinoma (OPSCC) is increasing in incidence despite a decline in traditional risk factors. Human Papilloma Virus (HPV), specifically subtypes 16, 18, 31 and 35, has been implicated as the high-risk etiologic agent.
10p vibaku2711 22-07-2020 10 1 Download
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The 16S rRNA gene is composed of highly conserved, specie-specific sequences between different species of bacteria and the application of restriction endonucleases on the amplified 16S rRNA gene is a novel diagnostic tool in molecular characterization of bacterial isolates.
10p nguaconbaynhay6 23-06-2020 11 0 Download
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Tomato leaf curl virus (ToLCV) is a major Geminivirus which cause serious loss to tomato production in tropical and subtropical regions of the world. Hence, considered as major constraint to tomato cultivation. In the present study ToLCV coat protein gene was cloned and expressed using recombinant DNA technology approach. The ToLCV infected tomato leaf samples were collected from tomato fields near Krishi Vigyan Kendra, Dharwad. Further, the total DNA from ToLCV infected tomato leaf sample was isolated by following CTAB protocol.
11p nguathienthan4 18-04-2020 11 2 Download
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A complex Mus81-Mm4 is a DNA structure–specific endonuclease in Saccharomyces cerevisiae. Mus81-Mms4 functions in processing of recombination intermediates that could arise during the repair of stalled and blocked replication forks and double stranded breaks. Mus81-Mms4 works with many proteins involved in DNA repair, replication fork stability, and joint molecule formation/resolution during homologous recombination repair.
5p vitunis2711 11-12-2019 11 0 Download
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In the present study, a novel pre-miniproinsulin analogue was designed to have a short 9 residue sequence replacing the 35 residue C-chain, one lysine and one arginine added to the C-terminus of the B-chain in combination with glycine and arginine substitution at A21 and B29, respectively, and a 16-residue fusion partner comprising the pentapeptide sequence (PSDKP) of the N-terminus of human tumor necrosis factor-a (TNF-a), 6 histidine residues for Ni2+ chelated affinity purification and a pentapeptide ending with methionine for ease of chemical cleavage fused at the N-terminus.
9p trinhthamhodang1 16-11-2019 23 1 Download
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The antibody titer and specificity were determined by ELISA and western blot analysis, respectively. The tissue distribution of pSix1 was determined by western blot using the prepared polyclonal antibody.
8p vikimsa 22-02-2019 18 1 Download
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Sequence analysis of aPaenibacillussp. BP-23 recombinant clone coding for a previously described endoglucanase revealed the presence of an additional truncated ORF with homology to family 48 glycosyl hydrolases. The corres-ponding 3509-bp DNA fragment was isolated after gene walking and cloned inEscherichia coliXl1-Blue for expres-sion and purification. The encoded enzyme, a cellulase of 1091 amino acids with a deduced molecular mass of 118 kDa and a pI of 4.
7p fptmusic 16-04-2013 49 3 Download
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Within this work we describe the purification and biochemical characteriza-tion of a ddNTP-sensitive DNA polymerase purified from mungbean (Vigna radiatacv B1, L.) seeds at 18 days after fertilization, when 70% of the nuclei are reported to be in the endoreduplicated state.
19p galaxyss3 19-03-2013 38 3 Download
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Three DNA regions carrying genes encoding putative homologs of xanthine dehydrogenases were identified inEscherichia coli, named xdhABC, xdhD, andyagTSRQ. Here, we describe the purification and characterization of gene products of the yagTSRQ operon, a molybdenum-containing iron– sulfur flavoprotein from E. coli, which is located in the periplasm.
13p viettel02 22-02-2013 16 2 Download