• Title/Summary/Keyword: DNA함량

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Antioxidant Properties in Microbial Fermentation Products of Lonicera japonica Thunb. Extract (금은화 추출물을 이용한 미생물 발효 생성물의 항산화 특성)

  • Shin, Jung-Hee;Yoo, Sun-Kyun
    • Journal of the East Asian Society of Dietary Life
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    • v.22 no.1
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    • pp.95-102
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    • 2012
  • The purpose of this study is to investigate antioxidant properties in microbial fermentation products of Lonicera japonica Thunb extract. The bacterium Lactobacillus plantarum NHP1 was isolated from conventional fermented foods. Modern pharmacological studies show that Lonicera japonica Thunb and its active principles of wide pharmacological actions. For instance, they show a strong efficacy in antibacterial, anti-inflammatory, antiviral, anti-endotoxin, blood fat reducing, antipyretic, and antioxidant activities. The extract of Lonicera japonica Thunb was obtained by extracting dried Lonicera japonica Thunb using either hot water or 70% ethanol as a solvent. Fermentation was performed in a 2L fermentor containing 1.2 L of extractat conditions of $30^{\circ}C$ and 100 rpm for 48 hr. The amount of cholorogenic acid was $2.65{\mu}g/g$ in hot water extract. The total phenolic content (GAE, gallic acid equivalent) in hot water and 70% ethanol were $56.5{\pm}4.9$ GAE mg/g and $72.7{\pm}5.3$ GAE mg/g, respectively. After fermentation, the phenolic content increased to 30.2% in hot water and 12.9% in ethanol extract. In the same manner, flavonoid content increased to more than 75% regardless of extract solvent. ABTS (2,2-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid) value noticeably increased to 50% after fermentation.

Breeding of Green Soybean Strain with Green Cotyledon and Tetra Null Genotype (Tetra null 유전자형과 녹색종피 및 자엽을 가진 콩 계통 육종)

  • Sarath Ly;Jeong Hwan Lee;Hyeon Su Oh;Se Yeong Kim;Jin Young Moon;Jong Il Chung
    • Journal of Life Science
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    • v.33 no.8
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    • pp.632-638
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    • 2023
  • A soybean cultivar with a green seed coat and cotyledon contains high levels of lutein, which is beneficial for eye health. Plus, antinutritional components such as lipoxygenase, Kunitz trypsin inhibitor (KTI), lectin and stachyose exist in the mature seed. The genetic elimination of these antinutritional factors is a necessary step in green soybean breeding. This research was conducted to improve a new green soybean line with the green cotyledon and tetra null genotype (lox1lox2lox3tilers2) in terms of lipoxygenase, KTI, lectin and stachyose. We used five germplasms to develop a breeding population. A total of 69 F2 seeds were obtained from the cross of parent 1 and parent 2, and from those, 21 F2 seeds were selected that had the green seed coat color, and which were free of lectin protein. Next, four F2 plants with the green seed coat and tetra null genotype were selected from the breeding population derived from four genotypes. The absence of lipoxygenase, KTI and lectin proteins was confirmed in the F5 strain. The breeding line has a green seed coat, green cotyledon and white hilum color. The 100-seed weight and stachyose content for the breeding line were 30.7 g and 2.40 g/kg, respectively. The line selected in this study could be used as a cultivar or parent to improve colored soybean cultivars through the removal of antinutritional components such as lip- oxygenase, KTI, lectin and stachyose.

Isolation and Identification of Oceanisphaera sp. JJM57 from Marine Red Algae Laurencia sp. (Ceramiales: Rhodomelaceae) (해양 홍조류 Laurencia sp. (Ceramiales: Rhodomelaceae)에서 분리한 Oceanisphaera sp. JJM57의 분리 및 동정)

  • Kim, Man-Chul;Dharaneedharan, S.;Moon, Young-Gun;Kim, Dong-Hwi;Son, Hong-Joo;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.58-63
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    • 2013
  • A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM $15406^T$ and Oceanisphera donghaensis KCTC $12522^T$, respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to $37^{\circ}C$. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2-OH, $C_{16:0}$, and $C_{18:1}$ ${\omega}7c$ fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

On the Secretion and Functions of Equine Chorionic Gonadotropin (말의 융모성 성선자극 호르몬의 분비와 기능)

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.2
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    • pp.125-142
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    • 2000
  • 13). Analysis of a purified preparation of eCG revealed that its $\beta$ -subunit consists of 149 amino acids, which was confirmed by the molecular cloning of its cDNA. There seem to be at least four to six, or even as many as 11, O-glycosylation sites on the extended C-tenninal region of the eCG $\beta$-subunit. Interestingly, eCG is a unique member of this family, as it appear to be a single molecule that possesses both LH- and FSH-like activities. Using the cDNA prepared from mRNA extracted from equine placental and pituitary tissues, we cloned the cDNA of eCG $\alpha$- and $\beta$ -subunits and eFSH $\beta$ -subunit. The mRNA expression of each subunit seems to be independently regulated, which may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. Thus, eCG is a distinct molecule from the view points of its biological function and glycoresidue structures. Recombinant eCGs including the mutants which lack oligosaccharides will be useful tools for analyzing the structure-function relationships of gonadotropins in the horse as well as other species. Similar experiments will also clarify the proposed structure and biological functions for the glycoprotein hormones. These experimental are now possible, and hopefully a resolution of the existing controversy will be forthcoming in the near future.

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Study on Characteristics of Chemical Properties and Microbial Flora of Organic Farming Soil in Korea (유기농 토양의 화학적 특성 및 미생물상 연구)

  • Park, Kwang-Lai;Suga, Yuko;Hong, Seung-Gil;Lee, Chorong;Ahn, Minsil;Kim, Seok-Cheol;Hashimoto, Tomoyoshi
    • Journal of the Korea Organic Resources Recycling Association
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    • v.24 no.4
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    • pp.77-83
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    • 2016
  • The objectives of this study was to investigate the difference between organic-farming and conventional-farming soils relatives to soil chemical properties and microbial flora. Fifteen soil sampling sites were chosen from the certified organic upland farm, considered with its location, crop and application of organic compost types. Soil chemical properties were analyzed by standard methods established by National Institute of Agricultural Sciences, Rural Development Administration. For the soil chemical properties, the values of pH were ranged from 4.5 to 7.3. The values of electrical conductivity (EC) in the sampling sites were below 2 dS/m of convention cultivation soil. For analyzing the microbial flora, the bacillus(16S rDNA) and cladothricosis(18S rDNA) were analyzed by using PCR-DGGE (Denaturing Gradient Gel Electrophoresis) in the soil of 15 sampling sites. Cluster analysis of biodiversity index was performed by using pattern of DGGE. DGGE patterns and clustering analysis of bacterial DNA from soil extracts revealed that the bacterial community was differentiated between less than 5 years and more than 5 years depending on the cultivation history. But there was no consistent tendency between cultivation history and regional trend in the case of molds. Therefore, it would be very effective to analyze bacterial clusters of organically cultivated soils in long - term cultivated soil for more than 5 years.

Isolation and Characterization of mas1+ of Schizosaccharomyces pombe, a Homologue of Human CIP29/Hcc-1 Involved in the Regulation of Cell Division (세포분열에 관여하는 인간의 CIP29/Hcc1 유전자와 상동성을 가지는 분열형 효모의 새로운 유전자 mas1+의 특성분석)

  • Cha, Jae-Young;Shin, Sang-Min;Ha, Se-Eun;Lee, Jung-Sup;Park, Jong-Kun
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1666-1677
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    • 2011
  • The regulation of gene expression plays an important role in cell cycle controls. In this study, a novel gene, the $mas1^+$($\underline{mi}$tosis $\underline{as}$sociated protein) gene, a homolog of human CIP29/Hcc1, was isolated and characterized from fission yeast Schizosaccharomyces pombe (S. pombe) using a gene-specific polymerase chain reaction. The isolated gene contained a complete open reading frame capable of encoding 245 amino acid residues with a typical promoter, as judged by nucleotide sequence analysis. It was also found that a PCB ($\underline{p}$ombe cell $\underline{c}$ycle $\underline{b}$ox) is located in the promoter region, which controls M-$G_1$ specific transcription in S. pombe. The quantitative analysis of the $mas1^+$ transcript against $adh1^+$ showed that the pattern of expression is similar to that of the septation index. Cytokinesis of mas1 mutant was greatly delayed at $25^{\circ}C$ and $36^{\circ}C$, and a large number of multi-septate cells were produced. The mas1 mutant had 2C, 4C and 6C DNA contents, as determined by FACS analysis. In addition, the number of multi-septate cells significantly increased. When cells were cultured in nitrogen starvation medium to increase proliferation, the abnormal phenotypes of mas1 mutant dramatically increased. These phenotypes could be rescued by an overexpression of the $mas1^+$ gene. The mas1 protein localized in the nuclei of S. pombe and human HeLa cells, as evidenced by Mas1-EGFP signals. The abnormal growth pattern and the morphology of mas1 mutant were complemented by a plasmid carrying human CIP29/Hcc-1cDNA. In addition, CIP29 /Hcc-1 transcript level increased in active cell proliferation stages in the developing mouse embryos. These results indicate that the $mas1^+$ ishomologous to the human CIP29/Hcc1 gene and is involved in cytokinesis and cell shape control.

Effects of Fermentation on the Metabolic Activities of Pine Needle Juice (발효과정이 솔잎 착즙액의 항산화, alpha-Glucosidase 및 Angiotensin Converting Enzyme 저해 활성에 미치는 영향)

  • Kim, So-Yun;Lee, Hyun-Jung;Park, Jae-Hee;Kim, Rae-Young;Cheong, Hyeonsook;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.3
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    • pp.325-334
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    • 2013
  • The objective of this study was to compare the content and metabolic activities between fresh pine needle juice (PNJ) and fermented pine needle juice (FPNJ). A variety of factors were measured, including total phenolic content (TPC), antioxidant activity [DPPH radical scavenging activity (RSA), total radical-trapping antioxidant potential (TRAP), oxygen radical absorbance capacity (ORAC), cellular antioxidant capacity (CAC)], anti-genotoxic activity, ${\alpha}$-glucosidase inhibitory activity, and angiotensin converting enzyme (ACE) inhibitory activity. The TPC was $17.3{\pm}0.2$ and $4.6{\pm}0.0$ mg GAE/g in PNJ and FPNJ, respectively. The DPPH RSA, TRAP, and ORAC values increased in a dose-dependent manner for both PNJ and FPNJ, with significantly higher activities in PNJ than FPNJ. The CAC against AAPH-induced oxidative stress in HepG2 cells was protected by both PNJ and FPNJ. Pretreatment with PNJ and FPNJ in human leukocytes produced significant reductions in $H_2O_2$-induced DNA damage at a concentration of $50{\mu}g/mL$. ${\alpha}$-Glucosidase inhibitory activity was significantly higher in FPNJ than PNJ. The ACE inhibitory activity was about 87.1% and 60.0% in 1:1 diluted PNJ and FPNJ, respectively. This study suggests that the fermentation of PNJ could enhance the regulation of blood glucose metabolism and both PNJ and FPNJ might be a new potential source of natural antioxidant, anti-diabetic, and anti-hypertensive agents applicable to food.

Isolation of Myrosinase and Glutathione S-transferase Genes and Transformation of These Genes to Develop Phenylethylisothiocyanate Enriching Chinese Cabbage (배추에서 항암물질 phenylethylisothiocyanate의 다량 합성을 위한 myrosinase와 glutathione S-transferase 유전자 분리 및 이를 이용한 형질전환체 육성)

  • Park, Ji-Hyun;Lee, Su-Jin;Kim, Bo-Ryung;Woo, Eun-Teak;Lee, Ji-Sun;Han, Eun-Hyang;Lee, Youn-Hyung;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.29 no.6
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    • pp.623-632
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    • 2011
  • To increase the anti-carcinogens phenylethylisothiocyanate (PEITC), myrosinase (MYR), and glutathione S-transferase (GST), genes related to PEITC pathway were isolated and the gene expressions were regulated by Agrobacterium transformation. Isolated cDNAs, MYR, and GST genes were 1,647 bp and 624 bp, respectively, and the protein expression was confirmed through pET system. Thereafter, we constructed a sense-oriented over-expressing myrosinase (pBMY) and RNAi down-regulated GST (pJJGST) binary vectors for the Chinese cabbage transformation. After the transformation, thirteen over-expressing transgenic Chinese cabbage plants (IMS) with pBMY and five down-regulated ones (IGA) with pJJGST were selected by PCR analysis. Selected $T_0$ transgenic plants were generated to $T_1$ plants by self-pollination. Based on the Southern blot analysis on these $T_1$ transgenic plants, 1-4 copies of T-DNA were transferred to Chinese cabbage genome. Thereafter, RNA expression level of myrosinase gene or GST gene was analyzed through real-time RT PCR of IMS, IGA, and non-transgenic inbred lines. In case of IMS lines, myrosinase gene was increased 1.03-4.25 fold and, in IGA lines, GST gene was decreased by 26.42-42.22 fold compared to non-transgenic ones, respectively. Analysis of PEITC concentrations using GC-MS it showed that some IMS lines and some IGA lines increased concentrations of PEITC up to 4.86 fold and up to 3.89 fold respectively compared to wild type. Finally in this study IMS 1, 3, 5, 12, and 15 and IGA 1, 2, and 4 were selected as developed transgenic lines with increasing quantities of anti-carcinogen PEITC.

Analysis of Genetic Relationship of Cordyceps militaris in Korea by Random Amplified Polymorphic DNA (한국산 번데기동충하초의 RAPD 분석에 의한 종내 그룹의 유전적 유연관계 분석)

  • Sung, Jae-Mo;Kim, Sang-Hee;Yoon, Chul-Sik;Sung, Gi-Ho;Kim, Yong-Wook
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.256-273
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    • 1999
  • Seventy two isolates of Cordyceps militaris collected from 11 sites in Korea, including two isolates from ATCC, were used to assess genetic variation within Cordyceps militaris. The anamorph stage and cultural characteristics of C. militaris were observed through microscope and investigated on PDA respectively. The anamorphs of C. militaris were identified to be Verticillium. Isolates of C. militaris showed different growth rates, morphology and color. Fifty six isolates of single ascospore and seventy two isolates of mass ascospore from C. militaris were analysed using by Random Amplified Polymorphic DNA (RAPD) for genetic relationship analysis. Fifty six single ascospore isolates fell into two groups by phenogram constructed from distance values using the UPGMA method in NTSYS-pc software: group A from artificial fruit body of C18 except for isolate 51; group B from artificial fruit body of C738. The average genetic distance value within group A is 0.150 and group B is 0.163. The average genetic distance value between the two groups is 0.221. The average genetic distance value within 56 single ascospores is 0.207 and 72 mass ascospores is 0.330. Genetic relationships were not found among 72 mass ascospore isolates obtained from eleven geographically distant populations.

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Antioxidative Activity of Rumex crispus L. Extract (소리쟁이 추출물의 항산화 효능)

  • Rhim, Tae-Jin;Choi, Moo-Young;Park, Hee-Juhn
    • Korean Journal of Plant Resources
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    • v.25 no.5
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    • pp.568-577
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    • 2012
  • The objective of this study was to investigate the antioxidative capacity of ethanol extracts from Rumex crispus L. The concentration of R. crispus L. extract at which DPPH radical scavenging activity was inhibited by 50% was 2.15 mg/mL, which was lower than that of ${\alpha}$-tocopherol (0.43 mg/mL), as compared to 100% by pyrogallol as a reference. Total antioxidant status was examined by total antioxidant capacity against ABTS radical reactions. Total antioxidant capacities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 0.47 and 2.33 mM Trolox equivalents, respectively, which were higher than those of ${\alpha}$-tocopherol. Superoxide scavenging activities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 21.5 and 78.9%, respectively, which were not significantly (p>0.05) different from those of catechin. Oxygen radical absorbance capacities of R. crispus L. extract at concentrations of 20 and 100 ${\mu}g/mL$ were 62.5 and 156.4 ${\mu}M$ Trolox equivalents, respectively, which were lower than those of ascorbic acid. Cupric reducing antioxidant capacities of R. crispus L. extract at concentrations of 0.1 and 1 mg/mL were 0.28 and 1.88 mM Trolox equivalents, which were similar or significantly (p<0.05) higher than those of ${\alpha}$-tocopherol, respectively. R. crispus L. extract prevented supercoiled DNA strand breakage induced by hydroxyl radical and peroxyl radical. Total phenolic contents of R. crispus L. extract at concentrations of 0.5 and 5 mg/mL were 0.58 and 3.85 mM gallic acid equivalents, respectively. R. crispus L. extract at concentration of 0.1 and 0.5 mg/mL inhibited 0.2 mM tert-butyl hydroperoxide-induced cytotoxicity by 38.5 and 63.5%, respectively, in HepG2 cell culture system. Thus, strong antioxidant and cytotoxicity-inhibiting effects of R. crispus L. extract seem to be due to, at least in part, the prevention from free radicals-induced oxidation as well as high levels in total phenolic contents.