• BMB Reports
• /
• v.32 no.6
• /
• pp.594-598
• /
• 1999

The Jun and Fos families of eukaryotic transcription factors form heterodimers capable of binding to their cognate DNA enhancer elements. We are interested in searching for inhibitors or antagonists of the binding of the Jun-Fos heterodimer to the activator protein-1 (AP-1) site. The basic-region leucine zipper (bZIP) domain of c-Fos was expressed as a fusion protein with glutathione S-transferase, and allowed to form a heterodimer with the bZIP domain of c-Jun. The heterodimer was bound to glutathione-agarose, to which were added radiolabeled AP-1 nucleotides. After thorough washing, the gel-bound radioactivity was counted. The assay is faster than the coventional electrophoretic mobility shift assay because the gel electrophoresis step and the autoradiography step are eliminated. Moreover, the assay is very sensitive, allowing the detection of picomolar quantities of nucleotides, and is not affected by up to 50% dimethylsulfoxide, a solvent for hydrophobic inhibitors. Curcumin and dihydroguaiaretic acid, recently known inhibitors of Jun-Fos-DNA complex formation, were applied to this Jun-GST-fused Fos system and revealed to decrease the dimer-DNA binding.

• Microbiology and Biotechnology Letters
• /
• v.13 no.1
• /
• pp.87-91
• /
• 1985

Hpa I endonuclease from Haemophilus parainfluenzae has been purified of homogeneity and its physical and ezymatic properties have been studied. For the purification of the enzyme, Heparin agarose, SP-sephadex C-25, DEAE-sephadex A-50 and phosphocellulose chromatography columns were used. The denatured and reduced form of the enzyme is a monomer of molecular weight of $30,000{\pm}1,000$ as judged by 10% polyacrylamide gel electrophoresis containing 0.1% sodium dodesyl sulfate. Hpa I endonuclease was maximally active at neutral pH (7.0 to 7.5) in the presence of 50 mM NaCl.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.18 no.6
• /
• pp.557-562
• /
• 1985

Moderately halophilic bacteria were collected from solar salt with Larsen medium containing $10\%$ NaCl. A total of 56 strains were isolated and tested for the presence of plasmid DNA by agarose gel electrophoresis. Twelve isolates ($21\%$) carried at least one kind of plasmid. Six different isolates among them were selected to study the molecular weight of plasmids and the morphological and physiological characters. Vibrio sp. 14, Alcaligenes sp. 63, Pseudomonas sp. 11, Flavobacterium sp. 38, Bacillus sp. 16, and Alcaligenes sp. 52 carried at least one plasmid of about 7.2 kbp, 6.4 kbp, 6.85 kbp, 8.5 kbp, 8.75 kbp, and 6.8 kbp respectively.

• The Korean Journal of Zoology
• /
• v.25 no.3
• /
• pp.123-129
• /
• 1982

In order to establish a biochemical genetic system in Drosophila populations in Korea, the $\\alpha$-glycerophosphate dehydrogenase alleles of eleven natural populations of D. melanogaster in Korea were examined by means of agarose gel electrophoresis. The results obtained are presented below: 1. $\\alpha$-Glycerophosphate dehydrogenase ($\\alpha$-GPDH) allele is scored for eleven natural populations of D. melanogaster in Korea, resulting that $\\alpha$-GPDH is found to be widely polymorphic for two electrophoretic variants. 2. The heterozygosity of $\\alpha$-GPDH is calculated as $40\\sim50%$. 3. The frequency of the FF genotype of $\\alpha$-GPDH is found to be roughly same as the SS genotype, but less than the FS genotype. 4. The F gene of $\\alpha$-GPDH is distributed almost frequently as the S gene.

• Microbiology and Biotechnology Letters
• /
• v.13 no.4
• /
• pp.345-348
• /
• 1985

Recombinant chimeric plasmid constructed with Xba I digested pUBl10 and -pE194 was transformed by polyethylene glycol induced protoplast transformation system into Bacillus subtilis BR 151 on the mannitol regeneration media, and two genes of antibiotics resistance were expressed simultaneously in the transfromant. Transformation frequency of the recombinant plasmid was 6.5 $\times$ 10$^{-5}$ on the mannitol regeneration agar plate containing neomycin and erythromycin. The replication of recombinant plasmid in the recipient cells was confirmed by the alkaline extraction method and agarose gel electrophoresis.

• Toxicological Research
• /
• v.13 no.1_2
• /
• pp.39-48
• /
• 1997

Cyclophosphamide(25, 50 or 100 mg/kg), orally administered to male Sprague-Dawley rats, caused a time- and dose-dependent thymic atrophy. In the light microscopic examination of the atrophic thymus, thymocytes with condensed or fragmented nucleus were multifocally observed in the cortical region, started to increase 8 hr after CPA treatment and reached to the maximal level at 16 hr, although such cells were not seen after 48 hr when the severe depletion of thymocytes were marked. In agarose gel electrophoresis to analyze the DNA changes, DNA extracted from atrophic thymus showed a oligonucleosomal laddering at the corresponding time to morphological changes. In an additional supportive experiment, thymocytes showing morphological changes, nuclear condensation or apoptotic body, exhibited a positive reaction to immunoperoxidase staining using in situ apoptosis detection kit. Separately, agarose gel electrophoresis of DNA from bone .marrow cells was performed to investigate the involvement of bone marrow cells in the process of thymocyte apoptosis. Although DNA laddering was slightly increased 2 and 4 hr after treatment, no clear correlation was inferred. Taken togather, it is concluded that thymocytes showing morphological changes in thymic atrophy induced by cyclophosphamide administration represent an apoptosis having biochemical nature of programmed cell death.

• Journal of Sericultural and Entomological Science
• /
• v.35 no.2
• /
• pp.120-128
• /
• 1993

The objective of this study is to identify plasmids of Bacillus thuringiensis var. kurstaki HD-1(B. t k HD-1) toxic to lepidopteran larvae. The results from agarose gel electrophoresis indicated that the bacterium contained 9 plasmids with approximate sizes of 1.4, 4.9, 5.4, 9.3, 10, 29, 44, 52, and 150 megadaltons(Md). By treating the wild type of B. t k HD-1 with either SDS or EtBr as curing agent, 26 cured mutants of the bacterium were obtained, 9 of them were crystallifereous(cry+) and the others acrystallifereous(cry-). Plasmids from B. t k HD-1 were transferred to B. cereus 569 strR cry- recipients(Bc569 M1). Among 13 isolates of Bc569 M1 transcipient, 11 of them were capable of producing the crystal toxic proteins. The plasmid patterns of Bc569 M1 transcipients and partially curved mutants of B. t k HD-1 on agarose gel electrophoresis suggested that the 29 and 44Md plasmids should be involved in the production of crystalline toxic proteins.

• Mycobiology
• /
• v.39 no.2
• /
• pp.96-102
• /
• 2011

We investigated diet supplementation with shiitake mushroom fruiting bodies on biochemical and histological changes in hypercholesterolemic rats. Six-wk old female Sprague-Dawley albino rats were divided into three groups of 10 rats each. A diet containing 5% Lentinus edodes fruiting bodies given to hypercholesterolemic rats reduced plasma total cholesterol, triglyceride, low-density lipoprotein (LDL), total lipid, phospholipids, and the LDL/high-density lipoprotein ratio by 34.33, 53.21, 75.00, 34.66, 25.73, and 71.43%, respectively. Feeding mushroom also significantly reduced body weight in hypercholesterolemic rats. However, it had no detrimental effects on plasma albumin, total bilirubin, direct bilirubin, creatinine, blood urea nitrogen, uric acid, glucose, total protein, calcium, sodium, potassium, chloride, inorganic phosphate, magnesium, or enzyme profiles. Feeding mushroom increased total lipid and cholesterol excretion in feces. The plasma lipoprotein fraction, separated by agarose gel electrophoresis, indicated that L. edodes significantly reduced plasma ${\beta}$ and pre-${\beta}$-lipoprotein but increased ${\alpha}$-lipoprotein. A histological study of hepatic cells by conventional hematoxylin-eosin and oil red-O staining showed normal findings for mushroom-fed hypercholesterolemic rats. These results suggest that shiitake mushrooms could be recommended as a natural cholesterol lowering substance in the diet.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.29 no.5
• /
• pp.843-848
• /
• 2000

The single cell-gel electrophoresis assay (comet assay) was used to identify irradiated beans. Soy beans, kidney beans, and red beans were irradiated with $^{60}Co$ gamma rays at 0.1, 0.3, 0.5, 0.7, and 1.0 kGy. Beans were peeled out, crushed lightly, and treated with phosphate-buffered saline (PBS) to extract cells. The extracted cell suspension was mixed with agarose gel solution and spread on an agarose precoated slide. After lysis of the cells, they were subjected to microgel electrophoresis for 2 minutes, and then silver-stained. We found that the DNA fragments of the irradiated samples were stretched, migrated out of the cells, and formed tails towards the anode giving the appearance of comets, while the unirradiated or the undamaged cells formed very short or no tails. The tail lengths of irradiated samples were significantly increased as irradiation dose increased at the above 0.3 kGy.

• The Plant Pathology Journal
• /
• v.34 no.6
• /
• pp.499-505
• /
• 2018

Huanglongbing (HLB, Citrus greening disease) is one of the most devastating diseases that threaten citrus production worldwide. Although HLB presents systemically, low titer and uneven distribution of these bacteria within infected plants can make reliable detection difficult. It was known loop-mediated isothermal amplification (LAMP) method has the advantages of being highly specific, rapid, efficient, and laborsaving for detection of plant pathogens. We developed a new LAMP method targeting gene contained tandem repeat for more rapid and sensitive detection of Candidatus Liberibacter asiaticus (CLas), putative causal agent of the citrus huanglongbing. This new LAMP method was 10 folds more sensitive than conventional PCR in detecting the HLB pathogen and similar to that of real-time PCR in visual detection assay by adding SYBR Green I to mixture and 1% agarose gel electrophoresis. Positive reactions were achieved in reaction temperature 57, 60 and $62^{\circ}C$ but not $65^{\circ}C$. Although this LAMP method was not more sensitive than real-time PCR, it does not require a thermocycler for amplification or agarose gel electrophoresis for resolution. Thus, we expect that this LAMP method shows strong promise as a reliable, rapid, and cost-effective method of detecting the CLas in citrus and can be applied for rapid diagnosis is needed.