• 한국환경성돌연변이발암원학회지
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• 제17권1호
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• pp.1-6
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• 1997

The RAD4 gene of Saccharomyces cerevisiae is essential for the incision step of UV-induced excision repair. A yeast RAD4 gene has been previously isolated by functional complementation. In order to identify the RAD4 homologous gene from fungus Coprinus cinereus, we have constructed cosmid libraries from electrophoretically separated chromosomes of the C. cinereus. The 13 C. cinereus chromosomes were resolved by pulse-field gel electrophoresis, hybridized with S. cerevisiae RAD4 DNA, and then isolated homologous C. cinereus chromosome. The insert DNA of the RAD4 homolog was contained 3.2 kb. Here, we report the partial cloning and characterization of fungus C. cinereus homolog of yeast RAD4 gene. Southern blot analysis confirmed that C. cinereus contains the sequence homologous DNA to RAD4 gene and this gene exists as a single copy in C. cinereus genome. When total RNA isolated from C. cinereus cells was hybridized with the 1.2 kb PvuII DNA fragment of the S. cerevisiae RAD4 gene, a 2.5 kb of transcript was detected. The level of the transcript did not increase upon UV-irradiation, suggesting that the RAD4 homologous gene in C. cinereus is not UV-inducible.

• 한국식품영양학회지
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• 제26권4호
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• pp.990-995
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• 2013

In the pursuit of practical usage for Codonopsis lanceolata and development of functional processed food material, the research on the product characterization is being performed through the manufacturing of Yanggaeng which is extracted from the Codonopsis lanceolata skin concentrate. By adding Codonopsis lanceolata skin extracts (0, 40, 60, 80, 100%) to the product, the chemical property characterization and organoleptic test of the produced Yanggaeng are being conducted. The scavenging effects on DPPH free radical have a significant proportional relationship with the addition of Codonopsis lanceolata skin extracts as 61.30 for 100% added group as compared with 52.72 for the null-addition group. It is considered that the antioxidative activity increases with the addition of the Codonopsis lanceolata skin extraction as the SOD similar activity rate was increased to 49.23% from 39.31%, the flavor preference was ranked as 80%>60%>0%>100%>40% showing that 60~80% of Codonopsis lanceolata skin extraction addition seems to be the proper amount.

• The Plant Pathology Journal
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• 제34권2호
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• pp.139-142
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• 2018

Pyrenophoratritici-repentis is the causal agent of tan spot. According to their ability to produce necrosis and/or chlorosis on a set of four differential bread wheats, the isolates of this fungus are currently grouped into eight races. When durum wheat genotypes were added to the differential set, a new virulence type was identified in Algeria. The isolates showing this virulence pattern are unable to attack bread wheat while they cause necrosis in durum genotypes. In this work, characterization of those isolates was based on pathological and molecular aspects. This included inoculation of bread and durum wheat, and virulence gene analysis using PCR and sequencing. The results showed that all isolates caused a resistance on all bread wheats of the differential set, while they produced necrosis in durum. ToxA and ToxB genes were amplified in all isolates, whereas toxb was absent. Sequence analysis for both genes showed no differences with those found in the two functional genes. The presence of two genes, ToxA and ToxB, despite the absence of symptoms usually caused by their products, suggests the existence of a new homologous for these two genes yet unknown. The presence of ToxA in the isolate unable to produce necrosis in Glenlea is reported for the first time.

• International Journal of Industrial Entomology and Biomaterials
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• 제21권2호
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• pp.157-162
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• 2010

Open reading frame 35 (bm35) of the Bombyx mori nucleopolyhedrovirus (BmNPV) is a special gene whose homologues are only found in some group-I nucleopolyhedroviruses, suggesting that bm35 plays a specific role in the viral life cycle. This paper described the characterization of BmNPV bm35. Computerassisted sequence analysis shows that a putative RING finger motif is observed in the protein, Bm35 encoded by bm35. The coding sequence of bm35 was amplified and subcloned into the vector pET30a(+) and the $(His)_6$-tagged fusion protein His-Bm35 was expressed in the Escherichia coli BL21 (DE3) LysS cells. The bm35 transcript and Bm35 protein were detected in BmNPV-infected BmN cells at 12~48 h post infection (p.i.) by RT-PCR and Western blot analysis using the polyclonal antibody generated by immunizing a rabbit with purified $(His)_6$-tagged Bm35, suggesting that bm35 is synthesized in the late stage of BmNPV infection cycle. Bm35 was not a structural component associated with budded virus (BV) and occlusion derived virus (ODV). These data indicated that bm35 is a functional gene in the BmNPV life cycle.

• Bulletin of the Korean Chemical Society
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• 제28권4호
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• pp.574-580
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• 2007

Full-length chloride intracellular channel protein 1 (CLIC1) is a member of the family of proteins related to bovine chloride intracellular channel p64. Mutations in the SEDL gene cause spondyloepiphyseal dysplasia tarda (SEDT), a rare X-linked chondrodysplasia. The link between the intracellular chloride channels and SEDL is an important step toward understanding their functional interplay. In the present study, CLIC1 protein was subcloned into the pGEX-KG vector and overexpressed in XL-1 blue cells. We developed a large-scale expression system composed of glutathione S-transferase (GST) fused with a 240-amino-acid CLIC1 protein in Escherichia coli. The soluble CLIC1 protein was successfully purified to homogeneity, and its purity, identity, activity and conformation were determined using SDS-PAGE, MALDI-MS, biophotometer and circular dichroism spectroscopic studies. The binding of both CLIC1 and SEDL proteins in vitro was detected by BIAcore biosensor and fluorescence measurements.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1692-1697
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• 2012

We report a glycoside hydrolase (GH)-118 ${\beta}$-agarase from a strain of Agarivorans, in which we previously reported recombinant expression and characterization of the GH-50 ${\beta}$-agarase. The GH comprised an open reading frame of 1,437 base pairs, which encoded a protein of 52,580 daltons consisting of 478 amino acid residues. Assessment of the entire sequence showed that the enzyme had 97% nucleotide and 99% amino acid sequence similarities to those of GH-118 ${\beta}$-agarase from Pseudoalteromonas sp. CY24, which belongs to a different order within the same class. The gene corresponding to a mature protein of 440 amino acids was inserted, recombinantly expressed in Escherichia coli, and purified to homogeneity with affinity chromatography. It had maximal activity at $35^{\circ}C$ and pH 7.0 and had 208.1 units/mg in the presence of 300 mM NaCl and 1 mM $CaCl_2$. More than 80% activity was maintained after 2 h exposure to $35^{\circ}C$; however, < 40% activity remained at $45^{\circ}C$. The enzyme hydrolyzed agarose to yield neoagarooctaose as the main product. This enzyme could be useful for industrial production of functional neoagarooligosaccharides.

• Archives of Pharmacal Research
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• 제29권7호
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• pp.605-616
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• 2006

A wide variety of drugs and endogenous bioactive amines are organic cations (OCs). Approximately 40% of all conventional drugs on the market are OCs. Thus, the transport of xenobiotics or endogenous OCs in the body has been a subject of considerable interest, since the discovery and cloning of a family of OC transporters, referred to as organic cation transporter (OCTs), and a new subfamily of OCTs, OCTNs, leading to the functional characterization of these transporters in various systems including oocytes and some cell lines. Organic cation transporters are critical in drug absorption, targeting, and disposition of a drug. In this review, the recent advances in the characterization of organic cation transporters and their distribution in the small intestine are discussed. The results of the in vitro transport studies of various OCs in the small intestine using techniques such as isolated brush-border membrane vesicles, Ussing chamber systems and Caco-2 cells are discussed, and in vivo knock-out animal studies are summarized. Such information is essential for predicting pharmacokinetics and pharmacodynamics and in the design and development of new cationic drugs. An understanding of the mechanisms that control the intestinal transport of OCs will clearly aid achieving desirable clinical outcomes.

• Journal of Microbiology and Biotechnology
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• 제31권5호
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• pp.696-704
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• 2021

Levan is an industrially important, functional biopolymer with considerable applications in the food and pharmaceutical fields owing to its safety and biocompatibility. Here, levan-type exopolysaccharide produced by Pantoea agglomerans ZMR7 was purified by cold ethanol precipitation and characterized using TLC, FTIR, ¹H, and ¹³C NMR spectroscopy. The maximum production of levan (28.4 g/l) was achieved when sucrose and ammonium chloride were used as carbon and nitrogen sources, respectively, at 35℃ and an initial pH of 8.0. Some biomedical applications of levan like antitumor, antiparasitic, and antioxidant activities were investigated in vitro. The results revealed the ability of levan at different concentrations to decrease the viability of rhabdomyosarcoma and breast cancer cells compared with untreated cancer cells. Levan appeared also to have high antiparasitic activity against the promastigote of Leishmania tropica. Furthermore, levan had strong DPPH radical scavenging (antioxidant) activity. These findings suggest that levan produced by P. agglomerans ZMR7 can serve as a natural biopolymer candidate for the pharmaceutical and medical fields.

• 한국식품영양학회지
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• 제34권3호
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• pp.302-309
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• 2021

This study examined the quality characteristics of yanggaeng with rice germ; it was incorporated into yanggaeng at different levels (containing 5% rice germ, 10% rice germ, 15% rice germ, and 20% rice germ) based on the total weight of red bean extracts. For analyzing the quality characteristics of yanggaeng, moisture content, hardness, color, antioxidant activity, total polyphenol content, reducing sugar, and vitamin E were determined. There was no significant difference in the moisture content and hardness except in the control. For color, lightness and yellowness of yanggaeng increased as the concentration of the powder was increased, whereas there was no significant difference in redness. As the rice germ powder was increased, total polyphenol content and antioxidant activity increased significantly, whereas reducing sugar decreased. Especially, total vitamin E, including isomers, increased as the concentration of the powder increased from 0.41 mg/100 g to 4.03 mg/100 g. Therefore, it could be possible to develop processed products with functional snack for yanggaeng prepared by adding 10% rice germ.

• Advances in nano research
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• 제11권6호
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• pp.657-665
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• 2021

The present work aimed to explore green approach via aqueous leaves extract of Murraya koenigii (ALEMk) for the synthesis of silver nanoparticles (AgNPsMk) in single step. The synthesis process was visualized with a color change and monitored by employing UV/Visible spectroscopy and a clear peak attained at 420 nm confirming the synthesis of AgNPsMk. The possible functional groups present in the extract which participated in the synthesis of AgNPsMk were identified with the help of FTIR spectroscopy. Further characterization using TEM images revealed the spherical shape of AgNPsMk with average particle size of 20 nm displaying well dispersion throughout the solution. Pronounced antioxidant activities of AgNPsMk at increased concentrations observed which evidencing strong radical scavenging ability. Moreover, AgNPsMk exhibited strong antibacterial behavior when tested against bacterial strains of Escherichia coli and Bacillus subtilis. Moving ahead, in vitro cytotoxicity work revealed potent cell viability loss appearing in AU565 and HeLa cancer cell lines on exposure to AgNPsMk at increased concentration. Finally, in vivo assessment carried out inside male Wistar rats indicated non toxic effect on examined liver tissues besides biochemical analysis including bilirubin, alkaline phosphtase (ALP) and serum glutamate pyruvate transaminase (SGPT) which found within the normal range when compared with control. The prior research work profoundly apprises the potential of green synthesized AgNPsMk to play a significant role in biomedical applications and formulations.