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Identification of the Differentially Expressed Genes of Hanwoo During the Growth Stage by Subtractive cDNA Hybridization (Subtraction 기법을 이용한 한우 성장 단계 특이 발현 유전자 탐색)

  • Jang, Y.S.;Kim, T.H.;Yoon, D.H.;Park, E.W.;Cheong, I.C.;Jo, J.K.
    • Journal of Animal Science and Technology
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    • v.44 no.1
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    • pp.13-22
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    • 2002
  • To identify the differentially expressed genes at growth stage of Hanwoo, we constructed the subtractive cDNA library from loin mRNA of 12- and 24-month old Hanwoo by PCR-based subtraction. The fourteen genes were confirmed by sequencing and reverse northern blot analysis, and they were selected as candidate of putative genes differentially expressed at the growth stage of Hanwoo. Three subtracted cDNA fragments that expressed specific signal to cDNA probe for 6-month-old loin of Hanwoo were highly homologous to those of the genes encoding EPV 20, Ca2+ATPase, and TCTP, respectively. The nine cDNA clones showed intense signal to cDNA probe from 12-month-old loin of Hanwoo, and highly homologus to those of genes encoding VCP, HSP 70, aldolase A, MSSK1, GM-2 activator protein, ryanodine receptor, acidic ribosomal phosphoprotein p1, ADP/ATP translocase, and UCP 2, respectively. Two subtracted cDNA clones that expressed specific signal to cDNA probes for 12- and 24-month-old loin of Hanwoo were detected. One of them was highly homologus to the gene encoding ferrochelatase and the other was highly homologus to the gene encoding ADRP.

Characterization of Caveola-Vesicle Complexes (CVCs) Protein, PHIST/CVC-8195 in Plasmodium vivax

  • Wang, Bo;Lu, Feng;Han, Jin-Hee;Lee, Seong-Kyun;Cheng, Yang;Nyunt, Myat Htut;Ha, Kwon-Soo;Hong, Seok-Ho;Park, Won Sun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • v.54 no.6
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    • pp.725-732
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    • 2016
  • Plasmodium vivax produces numerous caveola-vesicle complex (CVC) structures beneath the membrane of infected erythrocytes. Recently, a member helical interspersed subtelomeric (PHIST) superfamily protein, $PcyPHIST/CVC-81_{95}$, was identified as CVCs-associated protein in Plasmodium cynomolgi and essential for survival of this parasite. Very little information has been documented to date about $PHIST/CVC-81_{95}$ protein in P. vivax. In this study, the recombinant $PvPHIST/CVC-81_{95}$ N and C termini were expressed, and immunoreactivity was assessed using confirmed vivax malaria patients sera by protein microarray. The subcellular localization of $PvPHIST/CVC-81_{95}$ N and C termini in blood stage parasites was also determined. The antigenicity of recombinant $PvPHIST/CVC-81_{95}$ N and C terminal proteins were analyzed by using serum samples from the Republic of Korea. The results showed that immunoreactivities to these proteins had 61% and 43% sensitivity and 96.9% and 93.8% specificity, respectively. The N terminal of $PvPHIST/CVC-81_{95}$ which contains transmembrane domain and export motif (PEXEL; RxLxE/Q/D) produced CVCs location throughout the erythrocytic-stage parasites. However, no fluorescence was detected with antibodies against C terminal fragment of $PvPHIST/CVC-81_{95}$. These results suggest that the $PvPHIST/CVC-81_{95}$ is localized on the CVCs and may be immunogenic in natural infection of P. vivax.

Research article Black ginseng activates Akt signaling, thereby enhancing myoblast differentiation and myotube growth

  • Lee, Soo-Yeon;Go, Ga-Yeon;Vuong, Tuan Anh;Kim, Jee Won;Lee, Sullim;Jo, Ayoung;An, Jun Min;Kim, Su-Nam;Seo, Dong-Wan;Kim, Jin-Seok;Kim, Yong Kee;Kang, Jong-Sun;Lee, Sang-Jin;Bae, Gyu-Un
    • Journal of Ginseng Research
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    • v.42 no.1
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    • pp.116-121
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    • 2018
  • Background: Black ginseng (BG) has greatly enhanced pharmacological activities relative to white or red ginseng. However, the effect and molecular mechanism of BG on muscle growth has not yet been examined. In this study, we investigated whether BG could regulate myoblast differentiation and myotube hypertrophy. Methods: BG-treated C2C12 myoblasts were differentiated, followed by immunoblotting for myogenic regulators, immunostaining for a muscle marker, myosin heavy chain or immunoprecipitation analysis for myogenic transcription factors. Results: BG treatment of C2C12 cells resulted in the activation of Akt, thereby enhancing hetero-dimerization of MyoD and E proteins, which in turn promoted muscle-specific gene expression and myoblast differentiation. BG-treated myoblasts formed larger multinucleated myotubes with increased diameter and thickness, accompanied by enhanced Akt/mTOR/p70S6K activation. Furthermore, the BG treatment of human rhabdomyosarcoma cells restored myogenic differentiation. Conclusion: BG enhances myoblast differentiation and myotube hypertrophy by activating Akt/mTOR/p70S6k axis. Thus, our study demonstrates that BG has promising potential to treat or prevent muscle loss related to aging or other pathological conditions, such as diabetes.

A Study on the Daily Probability of Rainfall in the Taegu Area according to the Theory of Probaility (대구지방(大邱地方)의 확률일우량(確率日雨量)에 관(關)한 연구(硏究))

  • Kim, Young Ki;Na, In Yup
    • Economic and Environmental Geology
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    • v.4 no.4
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    • pp.225-234
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    • 1971
  • With the advance of civilization and steadily increasing population rivalry and competition for the use of the sewage, culverts, farm irrigation and control of various types of flood discharge have developed and will be come more and more keen in the future. The author has tried to calculated a formula that could adjust these conflicts and bring about proper solutions for many problems arising in connection with these conditions. The purpose of this study is to find out effective sewage, culvert, drainage, farm irrigation, flood discharge and other engineering needs in the Taegu area. If demands expand further a new formula will have to be calculated. For the above the author estimated methods of control for the probable expected rainfall using a formula based on data collected over a long period of time. The formula is determined on the basis of the maximum daily rainfall data from 1921 to 1971 in the Taegu area. 1. Iwai methods shows a highly significant correlation among the variations of Hazen, Thomas, Gumbel methods and logarithmic normal distribution. 2. This study obtained the following major formula: ${\log}(x-2.6)=0.241{\xi}+1.92049{\cdots}{\cdots}$(I.M) by using the relation $F(x)=\frac{1}{\sqrt{\pi}}{\int}_{-{\infty}}^{\xi}e^{-{\xi}^2}d{\xi}$. ${\xi}=a{\log}_{10}\(\frac{x+b}{x_0+b}\)$ ($-b<x<{\infty}$) ${\log}(x_0+b)=2.0448$ $\frac{1}{a}=\sqrt{\frac{2N}{N-1}}S_x=0.1954$. $b=\frac{1}{m}\sum\limits_{i=1}^{m}b_s=-2.6$ $S_x=\sqrt{\frac{1}{N}\sum\limits^N_{i=1}\{{\log}(x_i+b)\}^2-\{{\log}(x_0+b)\}^2}=0.169$ This formule may be advantageously applicable to the estimation of flood discharge, sewage, culverts and drainage in the Taegu area. Notation for general terms has been denoted by the following. Other notations for general terms was used as needed. $W_{(x)}$ : probability of occurranec, $W_{(x)}=\int_{x}^{\infty}f_{(n)}dx$ $S_{(x)}$ : probability of noneoccurrance. $S_{(x)}=\int_{-\infty}^{x}f_(x)dx=1-W_{(x)}$ T : Return period $T=\frac{1}{nW_{(x)}}$ or $T=\frac{1}{nS_{(x)}}$ $W_n$ : Hazen plot $W_n=\frac{2n-1}{2N}$ $F_n=1-W_x=1-\(\frac{2n-1}{2N}\)$ n : Number of observation (annual maximum series) P : Probability $P=\frac{N!}{{t!}(N-t)}F{_i}^{N-t}(1-F_i)^t$ $F_n$ : Thomas plot $F_n=\(1-\frac{n}{N+1}\)$ N : Total number of sample size $X_l$ : $X_s$ : maximum, minumum value of total number of sample size.

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Whole Genome Association Study to Detect Single Nucleotide Polymorphisms for Body Conformation Traits in a Hanwoo Population

  • Alama, M.;Lee, Y.M.;Park, B.L.;Kim, J.H.;Lee, S.S.;Shin, H.D.;Kim, K.S.;Kim, N.S.;Kim, J.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.3
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    • pp.322-329
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    • 2011
  • A whole genome association (WGA) study was conducted to identify quantitative trait loci (QTL) for body conformation traits in Hanwoo cattle. The phenotypes of 497 steers were recorded from the Hanwoo Improvement Center of National Agricultural Cooperative Federation, Seosan, Korea, and analyzed using the Illumina Bovine 50 k SNP chip. A set of 35,987 SNPs that were available in the Hanwoo population was selected from the chip. After adjustments for the effects of year-season of birth, region and sire, phenotypes were regressed on each SNP using a linear regression model. Three hundred nineteen SNPs were detected for the ten conformation traits (p<0.003). For the significant SNPs, stepwise regression procedures were applied to determine best sets of markers. A total of 72 SNPs were selected (p<0.001), for which the sets of 5, 9, 10, 9, 8, 11, 4, 6, 3 and 7 SNPs were determined for height at withers, rump height, body length, chest depth, chest width, rump length, hip width, thurl width, pinbone width and heart girth, respectively. About 7-26% of the total phenotypic variation was explained by the set of SNPs for each trait. QTL for the conformation traits were harbored on most bovine chromosomes (BTAs). Four SNPs with pleiotropic effects on height at withers and rump height were detected on BTAs 3, 4, 6 and 16. A SNP with pleiotropic effects on chest width and rump length was also detected on BTA10. Two QTL regions, i.e. between 87 and 97 Mb in BTA3 and between 41 and 44 Mb in BTA7, were found, in which SNPs were detected for the five and three conformation traits, respectively. The detected SNPs need to be validated in other Hanwoo populations for commercial application to the genetic improvement of conformation characteristics in Hanwoo via marker-assisted selection (MAS).

Functional Characterization of the ${\alpha}$- and ${\beta}$-Subunits of a Group II Chaperonin from Aeropyrum pernix K1

  • Lee, Jin-Woo;Kim, Se Won;Kim, Jeong-Hwan;Jeon, Sung-Jong;Kwon, Hyun-Ju;Kim, Byung-Woo;Nam, Soo-Wan
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.818-825
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    • 2013
  • We isolated and functionally characterized the ${\alpha}$- and ${\beta}$-subunits (ApCpnA and ApCpnB) of a chaperonin from Aeropyrum pernix K1. The constructed vectors pET3d-ApCpnA and pET21a-ApCpnB were transformed into E. coli Rosetta (DE3), BL21 (DE3), or CodonPlus (DE3) cells. The expression of ApCpnA (60.7 kDa) and ApCpnB (61.2 kDa) was confirmed by SDS-PAGE analysis. Recombinant ApCpnA and ApCpnB were purified by heat-shock treatment and anion-exchange chromatography. ApCpnA and ApCpnB were able to hydrolyze not only ATP, but also CTP, GTP, and UTP, albeit with different efficacies. Purified ApCpnA and ApCpnB showed the highest ATPase, CTPase, UTPase, and GTPase activities at $80^{\circ}C$. Furthermore, the addition of ApCpnA and ApCpnB effectively protected citrate synthase (CS) and alcohol dehydrogenase (ADH) from thermal aggregation and inactivation at $43^{\circ}C$ and $50^{\circ}C$, respectively. In particular, the addition of ATP or CTP to ApCpnA and ApCpnB resulted in the most effective prevention of thermal aggregation and inactivation of CS and ADH. The ATPase activity of the two chaperonin subunits was dependent on the salt concentration. Among the ions we examined, potassium ions were the most effective at enhancing the ATP hydrolysis activity of ApCpnA and ApCpnB.

Systematic approaches to identify functional genes using the FOX-hunting system in Chinese cabbage (FOX hunting system을 이용한 배추 기능유전자 탐색)

  • Lee, In-Hoo;Jung, Yu-Jin;Park, Jong-In;Nou, Ill-Sup;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.174-185
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    • 2010
  • Full-length cDNAs are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. To elucidate the functions of a large population of Chinese cabbage (Brassica rapa) genes and to search efficiently for agriculturally useful genes, we have been taking advantage of the full-length cDNA Over-eXpresser (FOX) gene hunting system. With oligo dT column it purify the each mRNA from the flower organs, leaf and stem tissue. And about 120,000 cDNAs from the library were transformed into $\lambda$-pFLCIII-F vector. Of which 115,000 cDNAs from the library were transformed into T-DNA binary vector, pBigs for transformation study. We used normalized full-length cDNA and introduced each cDNA into Arabidopsis by in planta transformation. Full-length Chinese cabbage cDNAs were expressed independently under the CaMV 35S promoter in Arabidopsis. Selfed seeds were harvested from transgenic Arabidopsis. We had selected 2,500 transgenic plants by hygromycin antibiotic tolerant test, and obtained a number of transgenic mutants. Each transgenic Arabidopsis was investigated in morphological changes, fertility and leaf colour. As a result, 285 possible morphological mutants were identified. Introduced cDNA was isolated by PCR amplification of the genomic DNA from the transgenic mutants. Sequencing result and BLAST analysis showed that most of the introduced cDNA were complete cDNAs and functional genes. Also, we examined the effect of Bromelain on enhancing resistance to soft rot in transgenic Chinese cabbage 'Osome'. The bromelain gene identified from FOX hunting system was transformed into Chinese cabbage using Agrobacterium methods. Transformants were screened by PCR, then RT-PCR and real time PCR were performed to analyze gene expression of cysteine protease in the T1 and T2 generations. The anti-bacterial activity of bromelain was tested in Chinese cabbages infected with soft rot bacteria. The results showed that the over-expressed bromelain gene from pineapple conferred enhanced resistance to soft rot in Chinese cabbage.

Estimating design floods for ungauged basins in the geum-river basin through regional flood frequency analysis using L-moments method (L-모멘트법을 이용한 지역홍수빈도분석을 통한 금강유역 미계측 유역의 설계홍수량 산정)

  • Lee, Jin-Young;Park, Dong-Hyeok;Shin, Ji-Yae;Kim, Tae-Woong
    • Journal of Korea Water Resources Association
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    • v.49 no.8
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    • pp.645-656
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    • 2016
  • The study performed a regional flood frequency analysis and proposed a regression equation to estimate design floods corresponding to return periods for ungauged basins in Geum-river basin. Five preliminary tests were employed to investigate hydrological independence and homogeneity of streamflow data, i.e. the lag-one autocorrelation test, time homogeneity test, Grubbs-Beck outlier test, discordancy measure test ($D_i$), and regional homogeneity measure (H). The test results showed that streamflow data were time-independent, discordant and homogeneous within the basin. Using five probability distributions (generalized extreme value (GEV), three-parameter log-normal (LN-III), Pearson type 3 (P-III), generalized logistic (GLO), generalized Pareto (GPA)), comparative regional flood frequency analyses were carried out for the region. Based on the L-moment ratio diagram, average weighted distance (AWD) and goodness-of-fit statistics ($Z^{DIST}$), the GLO distribution was selected as the best fit model for Geum-river basin. Using the GLO, a regression equation was developed for estimating regional design floods, and validated by comparing the estimated and observed streamflows at the Ganggyeong station.

Effect of Irradiation on the Quality Characteristics of Lyophilized Kimchi Powder (전자선 조사 동결건조 김치분말의 품질 특성)

  • Hong, S.P.;Kim, E.M.;Yang, J.N.;Ahn, D.U.
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.614-618
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    • 2007
  • The effect of irradiation on the quality characteristics of lyophilized kimchi powder was investigated in order to develop a commercial kimchi seasoning. Fresh and fermented kimchi powders were irradiated at 0, 1.5, 5, 10 and 30 kGy using a Linear Accelerator. By increasing the irradiation dose level, $a^*$ (redness) and $b^*$ (yellowness) values of the kimchi powders were decreased, while $L^*$ (lightness) value remained relatively unchanged as compared to the control. As the main volatile compounds, butanal, 2-butanone and acetic acid were produced in both of the kimchi powders at 30 kGy and dipropyl disulfide was detected only in the fermented kimchi. The viable counts of aerobic bacteria, yeasts, molds, and lactic acid bacteria in the kimchi powder were significantly reduced by all irradiation doses. However, aerobic and lactic acid bacteria were still observed in both of the kimchi powders at 30 kGy. No significant off-odors or off-tastes were produced in either of the kimchi powders by irradiation, while pungency decreased after irradiation. These results suggest that irradiated kimchi powder could be used as a kimchi seasoning.

Prediction of Chemical Composition and Fermentation Parameters in Forage Sorghum and Sudangrass Silage using Near Infrared Spectroscopy

  • Park, Hyung-Soo;Lee, Sang-Hoon;Choi, Ki-Choon;Kim, Ji-Hye;So, Min-Jeong;Kim, Hyeon-Seop
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.3
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    • pp.257-263
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    • 2015
  • This study was conducted to assess the potential of using NIRS to accurately determine the chemical composition and fermentation parameters in fresh coarse sorghum and sudangrass silage. Near Infrared Spectroscopy (NIRS) has been increasingly used as a rapid and accurate method to analyze the quality of cereals and dried animal forage. However, silage analysis by NIRS has a limitation in analyzing dried and ground samples in farm-scale applications because the fermentative products are lost during the drying process. Fresh coarse silage samples were scanned at 1 nm intervals over the wavelength range of 680~2500 nm, and the optical data were obtained as log 1/Reflectance (log 1/R). The spectral data were regressed, using partial least squares (PLS) multivariate analysis in conjunction with first and second order derivatization, with a scatter correction procedure (standard normal variate and detrend (SNV&D)) to reduce the effect of extraneous noise. The optimum calibrations were selected on the basis of minimizing the standard error of cross validation (SECV). The results of this study showed that NIRS predicted the chemical constituents with a high degree of accuracy (i.e. the correlation coefficient of cross validation ($R^2{_{cv}}$) ranged from 0.86~0.96), except for crude ash which had an $R^2{_{cv}}$ of 0.68. Comparison of the mathematical treatments for raw spectra showed that the second-order derivatization procedure produced the best result for all the treatments, except for neutral detergent fiber (NDF). The best mathematical treatment for moisture, acid detergent fiber (ADF), crude protein (CP) and pH was 2,16,16 respectively while the best mathematical treatment for crude ash, lactic acid and total acid was 2,8,8 respectively. The calibrations of fermentation products produced poorer calibrations (RPD < 2.5) with acetic and butyric acid. The pH, lactic acid and total acids were predicted with considerable accuracy at $R^2{_{cv}}$ 0.72~0.77. This study indicated that NIRS calibrations based on fresh coarse sorghum and sudangrass silage spectra have the capability of assessing the forage quality control