• Title/Summary/Keyword: Genomic effect

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Genomic DNA Methylation Status and Plasma Homocysteine in Choline- and Folate-Deficient Rats (콜린과 엽산 결핍이 흰쥐의 Genomic DNA 메틸화와 혈장 호모시스테인에 미치는 영향)

  • Mun, Ju-Ae;Min, Hye-Sun
    • Journal of Nutrition and Health
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    • v.40 no.1
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    • pp.14-23
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    • 2007
  • Elevated plasma homocysteine (Hcy) is a risk factor for cognitive dysfunction and Alzheimer disease, although the mechanism is still unknown. Both folate and betaine, a choline metabolite, play essential roles in the remethylation of Hcy to methionine. Choline deficiency may be associated with low folate status and high plasma Hcy. Alterations in DNA methylation also have established critical roles for methylation in development of the nervous system. This study was undertaken to assess the effect of choline and folate deficiency on Hcy metabolism and genomic DNA methylation status of the liver and brain. Groups of adult male Sprague Dawley rats were fed on a control, choline-deficient (CD), folate-deficient (FD) or choline/folate-deficient (CFD) diets for 8 weeks. FD resulted in a significantly lower hepatic folate (23%) (p<0.001) and brain folate (69%) (p<0.05) compared to the control group. However, plasma and brain folate remained unaltered by CD and hepatic folate reduced to 85% of the control by CD (p<0.05). Plasma Hcy was significantly increased by FD $(18.34{\pm}1.62{\mu}M)$ and CFD $(19.35{\pm}3.62{\mu}M)$ compared to the control $(6.29{\pm}0.60{\mu}M)$ (p<0.001), but remained unaltered by CD. FD depressed S-adenosylmethionine (SAM) by 59% (p<0.001) and elevated S-adenosylhomocysteine (SAM) by 47% in liver compared to the control group (p<0.001). In contrast, brain SAM levels remained unaltered in CD, FD and CFD rats. Genomic DNA methylation status was reduced by FD in liver (p<0.05) Genomic DNA hypomethylation was also observed in brain by CD, FD and CFD although it was not significantly different from the control group. Genomic DNA methylation status was correlated with folate stores in liver (r=-0.397, p<0.05) and brain (r = -0.390, p<0.05), respectively. In conclusion, our data demonsoated that genomic DNA methylation and SAM level were reduced by folate deficiency in liver, but not in brain, and correlated with folate concentration in the tissue. The fact that folate deficiency had differential effects on SAM, SAH and genomic DNA methylation in liver and brain suggests that the Hcy metabolism and DNA methylation are regulated in tissue-specific ways.

Effect of Sequence Variation in Bovine Mitochondrial DNA D-loop Region on Economic Traits for Hanwoo (한우 경제형질에 미치는 Mitochondrial DNA D-loop 영역의 염기서열 변이효과)

  • Oh, J.D.;Yoon, D.H.;Kong, H.S.;Lim, H.J.;Lee, H.K.;Cho, B.W.;Hong, K.C.;Jeon, G.J.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.933-938
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    • 2003
  • This study was performed to analyse the sequences of variations of mtDNA D-loop and their effects on carcass traits in Hnawoo(Korean cattle). The resulting sequences were compared with previously published sequences for other cattle breeds(GenBank J01394). The PCR was used to amplify a total of 964 bp between nucleotide 15758 and 383 within D-loop region of mtDNA using specific primers. Twenty five polymorphic sites by nucleotide substitution were found in mtDNA of Hanwoo. The frequencies of positions at 169, 16042, 16093, 16119, 16255 and 16302 nt with high levels of sequence polymorphism were 0.891, 0.117, 0.109, 0.182, 0.197 and 0.117, respectively. The substitution effect at 169 and 16119 nt was found significant on marbling score. Also substitution effect at 169 and 16042 nt was highly significant(p〈0.01) on backfat. thickness. Polymorphism of mtDNA sequence in D-loop region could be useful for the analysis of cytoplasmic genetic variation and associations with the other economically important traits and maternal lineage analysis in Hanwoo.

A Study on the Molecular Biology of Renin-Angiotensin System : Renin Gene Expression and Construction of Genomic Library (Renin-Angiotensin계의 분자생물학적 연구 : Renin유전자의 발현과 Genomic Library작성)

  • 박영순;한동민;김종호;문영희;이호섭;고건일;김성준
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.35-44
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    • 1990
  • Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.

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Chemoprevention of Helicobacter pylori-associated Gastric Carcinogenesis in a Mouse Model; Is It Possible?

  • Hahm, Ki-Baik;Song, Young-Joon;Oh, Tae-Young;Lee, Jeong-Sang;Surh, Young-Joon;Kim, Young-Bae;Yoo, Byung-Moo;Kim, Jin-Hong;Ha, Sang-Uk;Nahm, Ki-Taik;Kim, Myung-Wook;Kim, Dae-Yong;Cho, Sung-Won
    • BMB Reports
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    • v.36 no.1
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    • pp.82-94
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    • 2003
  • Although debates still exist whether Helicobacter pylori infection is really class I carcinogen or not, H. pylori has been known to provoke precancerous lesions like gastric adenoma and chronic atrophic gastritis with intestinal metaplasia as well as gastric cancer. Chronic persistent, uncontrolled gastric inflammations are possible basis for ensuing gastric carcinogenesis and H. pylori infection increased COX-2 expressions, which might be the one of the mechanisms leading to gastric cancer. To know the implication of long-term treatment of antiinflammatory drugs, rebamipide or nimesulide, on H. pylori-associated gastric carcinogenesis, we infected C57BL/6 mice with H. pylori, especially after MNU administration to promote carcinogenesis and the effects of the long-term administration of rebamipide or nimesulide were evaluated. C57BL/6 mice were sacrificed 50 weeks after H. pylori infection. Colonization rates of H. pylori, degree of gastric inflammation and other pathological changes including atrophic gastritis and metaplasia, serum levels and mRNA transcripts of various mouse cytokines and chemokines, and NF-${\kappa}B$ binding activities, and finally the presence of gastric adenocarcinoma were compared between H. pylori infected group (HP), and H. pylori infected group administered with long-term rebamipide containing pellet diets (HPR) or nimesulide mixed pellets (HPN). Gastric mucosal expressions of ICAM-1, HCAM, MMP, and transcriptional regulations of NF-${\kappa}B$ binding were all significantly decreased in HPR group than in HP group. Multi-probe RNase protection assay showed the significantly decreased mRNA levels of apoptosis related genes and various cytokines genes like IFN-$\gamma$, RANTES, TNF-$\alpha$, TNFR p75, IL-$1{\beta}$ in HPR group. In the experiment designed to provoke gastric cancer through MNU treatment with H. pylori infection, the incidence of gastric carcinoma was not changed between HP and HPR group, but significantly decreased in HPN group, suggesting the chemoprevention of H. pylori-associated gastric carcinogenesis by COX-2 inhibition. Long-term administration of antiinflammatory drugs should be considered in the treatment of H. pylori since they showed the molecular and biologic advantages with possible chemopreventive effect against H. pylori-associated gastric carcinogenesis. If the final concrete proof showing the causal relationship between H. pylori infection and gastric carcinogenesis could be obtained, that will shed new light on chemoprevention of gastric cancer, that is, that gastric/cancer could be prevented through either the eradication of H. pylori or lessening the inflammation provoked by H. pylori infection in high risk group.

The +1316 T/T Genotype in the Exon 3 of Uncoupling Protein Gene is Associated with Daily Percent Lay in Korean Native Chicken (한국 재래 닭의 Uncoupling Protein 유전자 Exon 3에서의 +1316 T/T 유전자형이 산란율에 미치는 효과 분석)

  • Oh J. D.;Lee J. H.;Hong Y. S.;Lee S. J.;Lee S. G.;Kong H. S.;Sang B. D.;Choi C. H.;Cho B. W.;Jeon G. J.;Lee H. K.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.239-244
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    • 2005
  • Uncoupling protein(UCP) is expressed exclusively in brown adipose tissue(BAT). It is blown to uncouple phosphorylation from oxidation and hence to be involved in energy metabolism and heat production, especially under cold exposure. In the present study, we identified single nucleotide polymorphism(SNP) in exon 3 of avUCP gene in Korean native chicken(KNC) population. It was detected a SNP T+1316C in exon 3 of avUCP gene by sequence analysis in KNC population. For PCR-RFLP analysis of the SNP T+1316C, used by AP III restriction enzyme. The result of PCR-RFLP analysis showed that allele T has two fragments of 255 bp and 86 bp, and allele C has only one fragment of 341 bp. The genotype frequencies were TT type, 0.7875; TC type, 0.1875 and CC type, 0.025; and the frequencies of allele T and C were 0.881 and 0.119, respectively in KNC population. Next study was conducted to investigate the effect of the SNP in avUCP gene on economic traits in the KNC population. The TT genotype had a significant higher daily percent lay(84.61) than CC genotype(p<0.05) in KNC population. This study may be useful for genetic studies of avCUP gene and selection on daily percent lay of KNC.

Prediction of genomic breeding values of carcass traits using whole genome SNP data in Hanwoo (Korean cattle) (한우에 있어서 유전체 육종가 추정)

  • Lee, Seung Hwan;Kim, Heong Cheul;Lim, Dajeong;Dang, Chang Gwan;Cho, Yong Min;Kim, Si Dong;Lee, Hak Kyo;Lee, Jun Heon;Yang, Boh Suk;Oh, Sung Jong;Hong, Seong Koo;Chang, Won Kyung
    • Korean Journal of Agricultural Science
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    • v.39 no.3
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    • pp.357-364
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    • 2012
  • Genomic breeding value (GEBV) has recently become available in the beef cattle industry. Genomic selection methods are exceptionally valuable for selecting traits, such as marbling, that are difficult to measure until later in life. One method to utilize information from sparse marker panels is the Bayesian model selection method with RJMCMC. The accuracy of prediction varies between a multiple SNP model with RJMCMC (0.47 to 0.73) and a least squares method (0.11 to 0.41) when using SNP information, while the accuracy of prediction increases in the multiple SNP (0.56 to 0.90) and least square methods (0.21 to 0.63) when including a polygenic effect. In the multiple SNP model with RJMCMC model selection method, the accuracy ($r^2$) of GEBV for marbling predicted based only on SNP effects was 0.47, while the $r^2$ of GEBV predicted by SNP plus polygenic effect was 0.56. The accuracies of GEBV predicted using only SNP information were 0.62, 0.68 and 0.73 for CWT, EMA and BF, respectively. However, when polygenic effects were included, the accuracies of GEBV were increased to 0.89, 0.90 and 0.89 for CWT, EMA and BF, respectively. Our data demonstrate that SNP information alone is missing genetic variation information that contributes to phenotypes for carcass traits, and that polygenic effects compensate genetic variation that whole genome SNP data do not explain. Overall, the multiple SNP model with the RJMCMC model selection method provides a better prediction of GEBV than does the least squares method (single marker regression).

Antioxidant and Nitric Oxide Inhibitory Activities of Pigments from Chionoecetes japonicas Rathbun (홍게 (Chionoecetes japonicas Rathbun) 껍질 색소의 항산화 활성 및 Nitric Oxide 생성억제 효과)

  • Park, Byungju;Baek, Seung Oh;Song, Young-Sun;Seo, Youngwan
    • KSBB Journal
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    • v.29 no.5
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    • pp.343-352
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    • 2014
  • In the present study, antioxidant activities of two crude pigments (acetone and MeOH) and their solvent fractions (n-hexane, 85% aq.MeOH, n-BuOH, and water fractions) from red crab shell were evaluated by measuring 1,1-diphenyl-2-picryl hydrazyl (DPPH), peroxynitrites, and degree of production of reactive oxygen species (ROS) in HT 1080 cells as well as the extent of oxidative damage of genomic DNA purified from HT 1080 cells. From comparative analysis, 85% aq.MeOH fraction showed the strongest scavenging effect on both peroxynitrite in vitro and intracellular ROS in HT 1080 cells. Protective activities of these samples against hydroxyl radical-mediated genomic DNA damage were also investigated. 85% aq.MeOH and n-BuOH fractions significantly inhibited oxidative damage of purified genomic DNA. On the other hand, we investigated their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. All samples significantly reduced NO production. Among the samples, n-hexane and water solvent fractions most effectively inhibited NO.

No excessive mutations in transcription activator-like effector nuclease-mediated α-1,3-galactosyltransferase knockout Yucatan miniature pigs

  • Choi, Kimyung;Shim, Joohyun;Ko, Nayoung;Park, Joonghoon
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.360-372
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    • 2020
  • Objective: Specific genomic sites can be recognized and permanently modified by genome editing. The discovery of endonucleases has advanced genome editing in pigs, attenuating xenograft rejection and cross-species disease transmission. However, off-target mutagenesis caused by these nucleases is a major barrier to putative clinical applications. Furthermore, off-target mutagenesis by genome editing has not yet been addressed in pigs. Methods: Here, we generated genetically inheritable α-1,3-galactosyltransferase (GGTA1) knockout Yucatan miniature pigs by combining transcription activator-like effector nuclease (TALEN) and nuclear transfer. For precise estimation of genomic mutations induced by TALEN in GGTA1 knockout pigs, we obtained the whole-genome sequence of the donor cells for use as an internal control genome. Results: In-depth whole-genome sequencing analysis demonstrated that TALEN-mediated GGTA1 knockout pigs had a comparable mutation rate to homologous recombination-treated pigs and wild-type strain controls. RNA sequencing analysis associated with genomic mutations revealed that TALEN-induced off-target mutations had no discernable effect on RNA transcript abundance. Conclusion: Therefore, TALEN appears to be a precise and safe tool for generating genomeedited pigs, and the TALEN-mediated GGTA1 knockout Yucatan miniature pigs produced in this study can serve as a safe and effective organ and tissue resource for clinical applications.

Inhibitory Effect of Genomic DNA Extracted from Pediococcus acidilactici on Porphyromonas gingivalis Lipopolysaccharide-Induced Inflammatory Responses

  • Young Hyeon Choi;Bong Sun Kim;Seok-Seong Kang
    • Food Science of Animal Resources
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    • v.43 no.1
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    • pp.101-112
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    • 2023
  • This study aimed to assess whether genomic DNA (gDNA) extracted from Pediococcus acidilactici inhibits Porphyromonas gingivalis lipopolysaccharide (LPS)-induced inflammatory responses in RAW 264.7 cells. Pretreatment with gDNA of P. acidilactici K10 or P. acidilactici HW01 for 15 h effectively inhibited P. gingivalis LPS-induced mRNA expression of interleukin (IL)-1β, IL-6, and monocyte chemoattractant protein (MCP)-1. Although both gDNAs did not dose-dependently inhibit P. gingivalis LPS-induced mRNA expression of IL-6 and MCP-1, they inhibited IL-1β mRNA expression in a dose-dependent manner. Moreover, pretreatment with both gDNAs inhibited the secretion of IL-1β, IL-6, and MCP-1. When RAW 264.7 cells were stimulated with P. gingivalis LPS alone, the phosphorylation of mitogen-activated protein kinases (MAPKs) was increased. However, the phosphorylation of MAPKs was reduced in the presence of gDNAs. Furthermore, both gDNAs restored IκBα degradation induced by P. gingivalis LPS, indicating that both gDNAs suppressed the activation of nuclear factor-κB (NF-κB). In summary, P. acidilactici gDNA could inhibit P. gingivalis LPS-induced inflammatory responses through the suppression of MAPKs and NF-κB, suggesting that P. acidilactici gDNA could be effective in preventing periodontitis.

Identification of a SNP in Chicken CaSR Gene and Its Effect on Economic Traits (닭의 CaSR 유전자내 단일 염기 변이 탐색 및 경제 형질간의 연관성 분석)

  • Hong, Y.S.;Oh, J.D.;Lee, J.H.;Kong, H.S.;Choi, C.H.;Lee, S.S.;Jeon, G.J.;Lee, H.K.
    • Korean Journal of Poultry Science
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    • v.34 no.2
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    • pp.151-156
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    • 2007
  • The Function of the calcium sensing receptor (CaSR) is to control calcium levels by altering PTH (parathyroid hormone) secretion and renal calcium resorption. The influence of calcium on the basal and stimulated release of several hormones from chicken pituitary glands has been determined in vitro. The objective of this study was to identify SNP in chicken CaSR gene and to investigate the effect of the SNP on economic traits. The sequencing analysis method was used to identify nucleotide polymorphisms within chicken CaSR gene. This study identified SNP at position 1949 bp(Genebank accession No : XM_416491) in the exon 1. The SNP changed the amino acid to alanine(GCC) from serine(TCC). This SNP showed three genotypes, AA, AS and SS by digestion with the restriction enzyme NcoⅠ using the PCR-RFLP method. The A963S showed significant effect only on the first lay day (P<0.05) in Leghorn population. Leghorn with the genotype AA had significantly faster the first lay day(137.6) than the genotype AS(143.0, P<0.05). Also, the A963S showed significant effect only on the first lay day(P<0.05) and mean of egg weight(P<0.05) in KNC population. KNC with the genotypes AA ans AS had significantly faster the first lay day (151.0 and 152.6, respectively) than the genotype SS(159.4, P<0.05). And the genotypes SS had significantly heavier the mean of egg weight(50.4 kg, P<0.05) than the genotype AA ans AS (47.5 and 47.8 kg, respectively). According to result of this study, an a allele of the A963S was found to have a significant effect on the first lay day. It will be possible to use this SNP marker on selecting chicken to improve the first lay day.