• Title/Summary/Keyword: ProteinChip

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Characterization and Gene Co-expression Network Analysis of a Salt Tolerance-related Gene, BrSSR, in Brassica rapa (배추에서 염 저항성 관련 유전자, BrSSR의 기능 검정 및 발현 네트워크 분석)

  • Yu, Jae-Gyeong;Lee, Gi-Ho;Park, Ji-Hyun;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.845-852
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    • 2014
  • Among various abiotic stress factors, soil salinity decreases the photosynthetic rate, growth, and yield of plants. Recently, many genes have been reported to enhance salt tolerance. The objective of this study was to characterize the Brassica rapa Salt Stress Resistance (BrSSR) gene, of which the function was unclear, although the full-length sequence was known. To characterize the role of BrSSR, a B. rapa Chinese cabbage inbred line ('CT001') was transformed with pSL94 vector containing the full length BrSSR cDNA. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that the expression of BrSSR in the transgenic line was 2.59-fold higher than that in the wild type. Analysis of phenotypic characteristics showed that plants overexpressing BrSSR were resistant to salinity stress and showed normal growth. Microarray analysis of BrSSR over-expressing plants confirmed that BrSSR was strongly associated with ERD15 (AT2G41430), a gene encoding a protein containing a PAM2 motif (AT4G14270), and GABA-T (AT3G22200), all of which have been associated with salt tolerance, in the co-expression network of genes related to salt stress. The results of this study indicate that BrSSR plays an important role in plant growth and tolerance to salinity.

Effects of Retinoic Acid on Differentiation and Gene Expression of Pig Preadipocytes (Retinoic Acid가 돼지 지방전구세포의 분화와 유전자 발현에 미치는 영향)

  • Lim, Hee-Kyong;Choi, Kang-Duk;Oyungerel, Baatartsogt;Choi, Young-Suk;Chung, Chung-Soo
    • Journal of Animal Science and Technology
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    • v.50 no.4
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    • pp.475-484
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    • 2008
  • The current study was undertaken to determine the effect of retinoic acid(RA) on differentiation and gene expression of pig preadipocytes. The preadipocytes were isolated from the backfat of the new-born pigs. RA was treated to the cultured cells for 4 days and RNA was extracted from the cells. Isolated RNA went through in situ hybridization using the 14,688-gene cDNA microarray chip. Degree of cell differentiation was determined by measuring glycerol 3-phosphate dehydrogenase activity. RA decreased differentiation of pig preadipocytes by 78%. Fourteen genes were significantly up-regulated by RA, including genes known to be involved in lipid metabolism, particulary sphingomyelin phosphodiesterase, apolipoprotein R precursor, growth factor receptor-bound protein 14, retinoic acid receptor RXR gamma. However, the expression of vascular endothelial growth factor D precursor and growth hormone receptor precursor genes playing a central role in cell growth, was greatly decreased. These results suggest that RA inhibits differentiation of pig preadiocytes by regulation of gene expression of the growth factor or growth hormone receptor.

Effects of Electrolyzed Alkaline Reduced Water on Echinostoma hortense Infection and Immune Response in C57BL/6 Mice (C57BL/6 생쥐에서 전해알칼리환원수가 호르텐스극구흡충 감염과 면역에 미치는 영향)

  • Kim, Dong-Heui;Deung, Young-Kun;Jin, Dan;Huang, Xue Zhu;Qi, Xu Feng;Kim, Kwang-Yong;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.38 no.1
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    • pp.11-19
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    • 2008
  • To examine the effect of the electrolyzed alkaline reduced water (ERW) on animal immunity, by employing Echinostoma hortense that is a parasite in the small intestine, the immune response of C57BL/6 was examined. To C57BL/6 mice, Echinostoma hortense metacercaria 15 per animal was in oculate dorsally, the worm was collected after 2 weeks, and the change of goblet cells and mast cells in the mucosa of small intestine was examined, and by using a protein chip, the change of cytokines in the serum was compared and observed. As a result, average 8.3 worms were collected from the C57BL/6 mice infected with E. hortense, and in the group fed with the ERW, average 10 worms were collected. In regard to the examination of the change of goblet cells, in the experimental group infected with E. hortense and fed with the ERW, average 4.3 worms per villus were detected, hence, it was found that the expression of goblet cells was low (p<0.001). Regarding the examination of the change of mast cells, similarly, in the group infected with E. hortense and fed with the ERW, average 11 worms per villus were detected, and it appears to be less than control group (p<0.001). Regarding the expression of cytokines in mouse serum, in comparison of the experiment group infected with E. hortense and control group, in the expression of the Th1 cytokines IL-6, IL-$1{\beta}$, IFN-${\gamma}$, TNF-${\alpha}$, and IL-2, and the Th2 cytokines IL-4, IL-5, IL-10, and IL-13, a significant difference was not detected. In our study, it was found that in the infection of E. hortense, the ERW mediates its effect on the number of goblet cells and mast cells in the intestinal mucosa, and simultaneously, the worm expulsion was delayed, and thus the conclusion that the ERW mediated its effect on the intestinal immunity of mice was obtained.

Identification of multiple key genes involved in pathogen defense and multi-stress tolerance using microarray and network analysis (Microarray와 Network 분석을 통한 병원균 및 스트레스 저항성 관련 주요 유전자의 대량 발굴)

  • Kim, Hyeongmin;Moon, Suyun;Lee, Jinsu;Bae, Wonsil;Won, Kyungho;Kim, Yoon-Kyeong;Kang, Kwon Kyoo;Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.43 no.3
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    • pp.347-358
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    • 2016
  • Brassinosteroid (BR), a plant steroid hormone, plays key roles in numerous growth and developmental processes as well as tolerance to both abiotic and biotic stress. To understand the biological networks involved in BR-mediated signaling pathways and stress tolerance, we performed comparative genome-wide transcriptome analysis of a constitutively activated BR bes1-D mutant with an Agilent Arabidopsis $4{\times}44K$ oligo chip. As a result, we newly identified 1,091 (562 up-regulated and 529 down-regulated) significant differentially expressed genes (DEGs). The combination of GO enrichment and protein network analysis revealed that stress-related processes, such as metabolism, development, abiotic/biotic stress, immunity, and defense, were critically linked to BR signaling pathways. Among the identified gene sets, we confirmed more than a 6-fold up-regulation of NB-ARC and FLS2 in bes1-D plants. However, some genes, including TIR1, TSA1 and OCP3, were down-regulated. Consistently, BR-activated plants showed higher tolerance to drought stress and pathogen infection compared to wild-type controls. In this study, we newly developed a useful, comprehensive method for large-scale identification of critical network and gene sets with global transcriptome analysis using a microarray. This study also showed that gain of function in the bes1-D gene can regulate the adaptive response of plants to various stressful conditions.

Effect of Carthami Tinctorii Fructus Herbal-acupuncture Solution(CTF-HAS) on Gene Expression in HepG2 carcinomar cells (Oligonucleotide chip를 이용한 홍화자약침액(紅花子藥鍼液)이 간암세포주(肝癌細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響))

  • Lee, Kyung-min;Lim, Seong-chul;Jung, Tae-young;Seo, Jung-chul;Han, Sang-won
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.215-225
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    • 2005
  • Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Oligonucleotide microarray approach were employed to screen the differential expression genes. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CTF-HAS(0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cytotoxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of CTF-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome U133 Plus 2.0., Affimatrix Co.). ResuIts : It has no cytotoxic effects on HepG2 cells in all concentrations (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$). More than twofold up-regulated genes were 19 genes. The number of more than twofold down-regulated genes was 13. Discussion : This study showed the screening of CTF-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

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Association of Clusterin Polymorphisms (-4453T<G, 5608T<C) with Coronary Heart Disease in Korean Population (한국인에서의 Clusterin의 유전자다형성(-4453T<G, 5608T<C)과 관상동맥질환과의 연관성)

  • Kim, Su-Won;Yoo, Min
    • Journal of Life Science
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    • v.20 no.4
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    • pp.584-588
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    • 2010
  • Clusterin is an 80 kDa heterodimetric glycosylated protein which plays diverse biological roles in various tissues and organs. Clusterin is reported to be associated with the pathogenesis of coronary artery disease and atherosclerosis. Therefore, we investigated the genotype for the T

QTL Scan for Meat Quality Traits Using High-density SNP Chip Analysis in Cross between Korean Native Pig and Yorkshire

  • Kim, S.W.;Li, X.P.;Lee, Y.M.;Choi, Y.I.;Cho, B.W.;Choi, B.H.;Kim, T.H.;Kim, J.J.;Kim, Kwan-Suk
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1184-1191
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    • 2011
  • We attempted to generate a linkage map using Illumina Porcine 60K SNP Beadchip genotypes of the $F_2$ offspring from Korean native pig (KNP) crossed with Yorkshire (YS) pig, and to identify quantitative trait loci (QTL) using the line-cross model. Among the genotype information of the 62,136 SNPs obtained from the high-density SNP analysis, 45,308 SNPs were used to select informative markers with allelic frequencies >0.7 between the KNP (n = 16) and YS (n = 8) F0 animals. Of the selected SNP markers, a final set of 500 SNPs with polymorphic information contents (PIC) values of >0.300 in the $F_2$ groups (n = 252) was used for detection of thirty meat quality-related QTL on chromosomes at the 5% significance level and 10 QTL at the 1% significance level. The QTL for crude protein were detected on SSC2, SSC3, SSC6, SSC9 and SSC12; for intramuscular fat and marbling on SSC2, SSC8, SSC12, SSC14 and SSC18; meat color measurements on SSC1, SSC3, SSC4, SSC5, SSC6, SSC10, SSC11, SSC12, SSC16 and SSC18; water content related measurements in pork were detected on SSC4, SSC6, SSC7, SSC10, SSC12 and SSC14. Additional QTL of pork quality traits such as texture, tenderness and pH were detected on SSC6, SSC12, SSC13 and SSC16. The most important chromosomal region of superior pork quality in KNP compared to YS was identified on SSC12. Our results demonstrated that a QTL linkage map of the $F_2$ design in the pig breed can be generated with a selected data set of high density SNP genotypes. The QTL regions detected in this study will provide useful information for identifying genetic factors related to better pork quality in KNP.

Effects of 5-azacytidine, a DNA methylation inhibitor, on embryogenic callus formation and shoot regeneration from rice mature seeds (벼 성숙종자로부터 배상체 캘러스 형성 및 식물체 재분화에 DNA methylation 억제제인 5-azacytidine의 영향)

  • Lee, Yeon-Hee;Lee, Jung-Sook;Kim, Soo-Yun;Sohn, Seong-Han;Kim, Dool-Yi;Yoon, In-Sun;Kweon, Soon-Jong;Suh, Seok-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.133-140
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    • 2008
  • The modification of DNA and histone plays an important role for gene expression in plant development. The objective of this research is to observe the effects of methylation on the gene expression during dedifferentiation from rice mature seeds to callus and differentiation from callus to shoots. The embryogenic callus with ability to shoot regeneration was not induced on the N6A medium supplemented with 5-azacytidine and abnormal callus with brown color was formed. When the normal rice callus was placed on the regeneration MSRA medium supplemented with 5-azacytidine, the shoot regeneration was inhibited. The results showed that 5-azacytidine, DNA demethylating agent, had negative effects on normal embryogenic callus formation and shoot regeneration. This suggested that DNA methylation of some genes was required for normal cell dedifferentiation and differentiation in tissue culture. The microarray and $GeneFishig^{TM}$ DEG screening were used to observe the gene transcript profile in callus induction and regeneration on N6A (N6 medium + 5-azaC) and MSRA (MS regeneration medium + 5-azaC). Subsets of genes were up-regulated or down-regulated in response to 5-azaC treatments. The genes related with epigenetic regulation, electron transport, nucleic acid metabolism and response to stress were up and down regulated. The different expression of some genes (germin like protein etc.) during callus induction and shoot regeneration was confirmed using RT-PCR and northern blot analysis.

Assessment of Relationship between Fyn-related Kinase Gene Polymorphisms and Overweight/Obesity in Korean Population

  • Jung, Mi-Young;Kim, Bum-Shik;Kim, Youn-Jung;Koh, In-Song;Chung, Joo-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.2
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    • pp.83-87
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    • 2008
  • The fyn-related kinase (FRK) belongs to the tyrosine kinase family of protein kinases. Recent studies have shown that Frk affects pancreatic beta cell number during embryogenesis and promotes beta cell cytotoxic signals in response to streptozotocin. To investigate the genetic association between FRK polymorphisms and the risk of obesity in Korean population, single nucleotide polymorphisms (SNPs) in the FRK gene region were selected and analyzed. The body mass index (BMI) was calculated, and biochemical data (systolic blood pressure, diastolic blood pressure, hemoglobin A1C, triglyceride, total cholesterol, high density lipoprotein, and low density lipoprotein) of blood sample from each subject were also measured. One hundred fifty five healthy control and 204 overweight/obesity subjects were recruited. Genotype frequencies of six SNPs [rs6568920 (+8391G>A), rs3756772 (+56780A>G), rs3798234 (+75687C>T), rs9384970 (+68506G>A), rs1933739 (+72978G>A), and rs9400883 (+75809A>G)] in the FRK gene were determined by Affymetrix Targeted Genotyping Chip data. According to the classification of Korean Society for the Study of Obesity, control (BMI 18 to < 23) and overweight/obesity (BMI$\geq$23) subjects were recruited. For the analysis of genetic data, EM algorithm, SNPStats, Haploview, HapAnalyzer, SNPAnalyzer, and Helixtree programs were used. Multiple logistic regression analysis (codominant, dominant, and recessive models) was performed. Age and gender as covariates were adjusted. For biochemical data, Student's t test was used. The mean value of BMI in the control and overweigh/obesity groups was 21.1${\pm}$1.2 (mean${\pm}$SD) and 25.6${\pm}$2.0, respectively. All biochemical data of the overweight/obesity group were statistically significance, compared with the control group. Among six SNPs, two linkage disequilibrium (LD) blocks were discovered. One block consisted of rs1933739 and rs9400883, and the other comprised rs3756772 and rs3798234. One SNP (rs9384970, +68506G>A) showed an association with overweight/obesity in the codominant model (p=0.03). Interestingly, the AA genotype distribution in the overweight/obesity group (n=7, 3.5%) was higher than those in the control group (n=1, 0.6%), which is not found in either Japanese or Chinese subjects. Therefore, the AA genotype of rs9384970 may be a risk factor for development of obesity in Korean population. The results suggest that FRK may be associated with overweight/obesity in Korean population.

Isolation and Functional Identification of BrDSR, a New Gene Related to Drought Tolerance Derived from Brassica rapa (배추 유래 신규 건조 저항성 관련 유전자, BrDSR의 분리 및 기능 검정)

  • Yu, Jae-Gyeong;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.33 no.4
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    • pp.575-584
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    • 2015
  • Drought stress is a crucial environmental factor determining crop survival and productivity. The goal of this study was to clearly identify a new drought stress-tolerance gene in Brassica rapa. From KBGP-24K microarray data with the B. rapa ssp. pekinensis inbred line 'Chiifu' under drought stress treatment, a gene which was named BrDSR (B. rapa Drought Stress Resistance) was chosen among 738 drought-responsive unigenes. BrDSR function has yet to be determined, but its expression was induced over 6-fold by drought. To characterize BrDSR, the gene was isolated from B. rapa inbred line 'CT001' and found to contain a 438-bp open reading frame encoding a 145 amino acid protein. The full-length cDNA of BrDSR was used to construct an over-expression vector, 'pSL100'. Tobacco transformation was then conducted to analyze whether the BrDSR gene can increase drought tolerance in plants. The BrDSR expression level in T1 transgenic tobacco plants selected via PCR and DNA blot analyses was up to 2.6-fold higher than non-transgenic tobacco. Analysis of phenotype clearly showed that BrDSR-expressing tobacco plants exhibited more tolerance than wild type under 10 d drought stress. Taking all of these findings together, we expect that BrDSR functions effectively in plant growth and survival of drought stress conditions.