• Biomolecules & Therapeutics
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• v.6 no.1
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• pp.89-94
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• 1998

CJ50001 is a recombinant human granulocyte colony-stimulating facto, (rHuG-CSF) that stimulates the formation of neutrophils from bone marrow stem cells. It was produced in E. colt and purified through refolding and several processes. We produced CS970125(300) using purified C150001 and additives in order to test the stability of CJ50001. When CS970125(300) was stored at 50'S for more than 1 week, high molecular weight proteins were formed and those proteins were detected by non-reducing SDS-PAGE, gel filtration HPLC, and Western blot. Those proteins showed single band at the same position of CJ50001 in reducing SDS-PAGE. These data indicated that those high molecular weight proteins were the multimers of C150001. In biological assays, iu viro and in viro, the multimers did not have biological activity and inhibitory action to that of CJ 50001. The mutimers did not induce toxicity in mice and rats in acute toxicity test. These results suggest that if Cs970125(300) containing CJ50001 is stored at 5$0^{\circ}C$, CJ50001 will be the multimers that do not have biological activity and inhibitory effect to CJ50001 and do not induce acute toxicity.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.4
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• pp.259-266
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• 2019

Emulsion polymerase chain reaction (PCR) is performed on compartmentalized DNA, allowing a large number of PCR reactions to be carried out in parallel. Emulsion PCR has unique advantages in DNA amplification. It can be applied in many molecular biological assays, especially those requiring highly sensitive and specific DNA amplification. This review discusses the principle of emulsion PCR and its applications in biotechnology. Related technologies are also discussed.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.04a
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• pp.50.2-50
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• 1999

• BMB Reports
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• v.41 no.8
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• pp.581-586
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• 2008

The pre-replication complex (pre-RC), including the core hexameric MCM2-7 complex, ensures that the eukaryotic genome is replicated only once per cell division cycle. In this study, we identified a rice $\underline{m}ini\underline{c}hromosome$ $\underline{m}aintenance$ (MCM) homologue (OsMCM2) that functionally complemented fission yeast MCM2 (CDC19) mutants. We found OsMCM2 transcript expression in roots, leaves, and seeds, although expression levels differed slightly among the organs. Likewise, the OsMCM2 protein was ubiquitously expressed, but it was downregulated when nutritients were limiting, indicating that MCM2 expression (and therefore cell cycle progression) requires adequate nutrition. Yeast two-hybrid and GST pull-down assays demonstrated that OsMCM2 interacted with the COP9 signalosome 5 (CSN5). Taken as a whole, our results indicated that OsMCM2 functions as a subunit of the rice MCM complex and interacts with CSN5 during developmental regulation.

• Journal of Microbiology and Biotechnology
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• v.29 no.5
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• pp.713-720
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• 2019

Acanthamoeba castellanii belonging to the T4 genotype may cause a fatal brain infection known as granulomatous amoebic encephalitis, and the vision-threatening eye infection Acanthamoeba keratitis. The aim of this study was to evaluate the antiamoebic effects of three clinically available antidiabetic drugs, Glimepiride, Vildagliptin and Repaglinide, against A. castellanii belonging to the T4 genotype. Furthermore, we attempted to conjugate these drugs with silver nanoparticles (AgNPs) to enhance their antiamoebic effects. Amoebicidal, encystation, excystation, and host cell cytotoxicity assays were performed to unravel any antiacanthamoebic effects. Vildagliptin conjugated silver nanoparticles (Vgt-AgNPs) characterized by spectroscopic techniques and atomic force microscopy were synthesized. All three drugs showed antiamoebic effects against A. castellanii and significantly blocked the encystation. These drugs also showed significant cysticidal effects and reduced host cell cytotoxicity caused by A. castellanii. Moreover, Vildagliptin-coated silver nanoparticles were successfully synthesized and are shown to enhance its antiacanthamoebic potency at significantly reduced concentration. The repurposed application of the tested antidiabetic drugs and their nanoparticles against free-living amoeba such as Acanthamoeba castellanii described here is a novel outcome that holds tremendous potential for future applications against devastating infection.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.5
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• pp.2081-2086
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• 2015

Gecko is a kind of traditional Chinese medicine with remarkable antineoplastic activity. However, undefined mechanisms and ambiguity regarding active ingredients limit new drug development from gecko. This study was conducted to assess anti-angiogenic properties of the aqueous extracts of fresh gecko (AG) or macromolecular components separated from AG (M-AG). An enzyme-linked immunosorbent assay (ELISA) approach was applied to detect the vascular endothelial growth factor (VEGF) secretion of the tumor cells treated with AG or M-AG. The effect of AG or M-AG on vascular endothelial cell proliferation and migratory ability was analyzed by tetrazolium dye colorimetric method, transwell and wound-healing assays. Chick embryo chorioallantoic membrane (CAM) assays were used to ensure the anti-angiogenic activity of M-AG in vivo. The results showed that AG or M-AG inhibited the VEGF secretion of tumor cells, the relative inhibition rates of AG and M-AG being 27.2% and 53.2% respectively at a concentration of $20{\mu}L/mL$. AG and M-AG inhibited the vascular endothelial (VE) cell proliferation with IC50 values of $11.5{\pm}0.5{\mu}L/mL$ and $12.9{\pm}0.4{\mu}L/mL$ respectively. The VE cell migration potential was inhibited significantly (p<0.01) by the AG (${\geq}24{\mu}L/mL$) or M-AG (${\geq}12\mu}L/mL$) treatment. In vivo, neovascularization of CAM treated with M-AG was inhibited significantly (p<0.05) at a concentration of ${\geq}0.4{\mu}L/mL$. This study provided evidence that anti-angiogenesis is one of the anti-tumor mechanisms of AG and M-AG, with the latter as a promising active component.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.3
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• pp.295-301
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• 2012

This study was carried out to determine the biological activities of Wa-song (Orostachys japonicus) hot water extracts. Four types of extract samples were prepared, including Wa-song, traditional plants mixture [PM; mixture of Baekbokryung (Poria cocos), Changchul (Atractylodis rhizoma), and Sa-in (Amomum xanthoides)], and two different ratio composites of these (mixture of PM and Wa-song extract, 1:1 (v/v); PMO-1 and 1:3 (v/v); PMO-3). Their biological activities were measured using various in vitro assays. Total phenolic and flavonoid contents of PM were higher compared to those of Wa-song, and those of PMO-1 were higher than those of PMO-3. Further, PMO-1 contained higher ABTS and DPPH radical scavenging, reducing power, and nitrite scavenging activities than PMO-3. On the contrary, PMO-3 contained higher tyrosinase and inhibitory activities of MCF-7 and HT-29 cancer cells than PMO-1. According to the results, biological activities of PMOs were significantly higher than those of Wa-song extract and PM in in vitro assays. Therefore, we expect that PMOs could show higher biological activities than Wa-song extract alone in vivo.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.896-900
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• 2006

Cytokinin has been known to act as a plant hormone to promote cell division and function in diverse processes in plant growth and development. Besides being produced in plants, it is also produced by various bacteria and fungi; however, its ecological significance is still unclear. In this report, we present an interesting finding that transzeatin riboside (tZR), a naturally occurring cytokinin compound, increased antibiotic production in many different streptomycetes, including Streptomyces coelicolor Ml3O, S. pristinaespiralis ATCC 25486, S. violaceoruber Tu22, S. anfibioticus ATCC l1891, and S. griseus IFO 13350. In vitro plate assays showed that the addition of 100 $\mu$M tZR increased the growth inhibition of Pseudomonas syringae pv. syringae, a plant pathogen, by S. griseus, a streptomycin producer. We suggest that cytokinin could act as a signaling molecule for antibiotic production in streptomycetes, a group of rhizosphere bacteria.

• Mycobiology
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• v.48 no.1
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• pp.51-57
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• 2020

A polysaccharide (LVP) was purified from fruiting body of Lentinus velutinus by ethanol precipitation fractionation and DEAE and Sephadex G-100 column chromatography. The yield of purified polysaccharide was 0.025%. Molecular characteristics of LVP were determined by gel permeation chromatography, FT-IR spectroscopy, and thin-layer chromatography. Our results revealed that LVP is a polysaccharide composed of only glucose units, and has a molecular weight of 336 kDa. Biological activity assays indicated that LVP exhibits both cytotoxic and antioxidant activity. LVP showed specific cytotoxicity against cancer cells (HeLa and HepG2 cells), and alterations in cancer cell morphology were found after LVP treatment.

• BMB Reports
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• v.38 no.6
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• pp.690-694
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• 2005

Transcription termination of the human mitochondrial genome requires specific binding to termination factor mTERF. In this study, mTERF was produced in E. coli and purified by two-step chromatography. mTERF-binding DNA sequences were isolated from a pool of randomized sequences by the repeated selection of bound sequences by gel-mobility shift assay and polymerase chain reaction. Sequencing and comparison of the 23 isolated clones revealed a 16-bp consensus sequence of 5'-GTG$\b{TGGC}$AGANCCNGG-3' in the light-strand (underlined residues were absolutely conserved), which nicely matched the genomic 13-bp terminator sequence 5'-$\b{TGGC}$AGAGCCCGG-3'. Moreover, mTERF binding assays of heteroduplex and single-stranded DNAs showed mTERF recognized the light strand in preference to the heavy strand. The preferential binding of mTERF with the light-strand may explain its distinct orientation-dependent termination activity.