• 제목/요약/키워드: PCR (polymerase chain reaction)

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돼지 Melanocortin 4 Receptor (MC4R) 유전자의 육질연관성 분석 (Characterization and Evaluation of Melanocortin 4 Receptor (MC4R) Gene Effect on Pork Quality Traits in Pigs)

  • 노정건;김상욱;최정석;최양일;김종주;최봉환;김태헌;김관석
    • Journal of Animal Science and Technology
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    • 제54권1호
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    • pp.1-8
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    • 2012
  • 본 연구는 한국재래돼지의 MC4R 유전자 내의 단일염기변이들을 규명하고 그 유전자형 효과가 유전자표지인자를 이용한 선발(Marker assisted selection, MAS)에 활용 가능한지를 검증하기 위해서 수행되었다. 한국재래돼지의 MC4R 유전자 총 염기서열을 분석하기 위해 6개의 Primer들을 이용하여 증폭산물을 생성하였으며, 염기서열분석을 통해 총 6개(c.-780C>G, c-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, c.*430A>T)의 단일염기변이를 발견하였다. 한국재래돼지 MC4R 유전자내의 총 6개의 단일염기변이들간의 연관불균형과 반수체 분석을 통해 단일염기변이들간의 연관성을 분석하였으며, c.-780C>G, c-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His와 c.*430A>T는 완전한 연관불균형을 이루고 있었고, c.892A>G(Asp298Asn) 단일염기변이만 $r^2$-value가 0.028, D'-value가 0.348로 연관불균형 정도가 매우 낮았다. c.707A>G (Arg236His)와 c.892A>G (Asp298Asn) 단일염기변이들을 선발하여 PCR-RFLP 유전자형 분석방법을 이용해 돼지 5품종간의 유전자형 빈도를 추정한 결과, c.707A>G (Arg236His) 단일염기변이는 요크셔 품종 집단에서 오직 A (His) 대립유전자를 관찰할 수 있었으며, 나머지 한국재래돼지, 랜드레이스, 버크셔와 듀록 품종에서는 G 대립유전자의 고정으로 나타났다. c.707A>G 단일염기변이와 육질형질을 484두에서 연관성 분석을 실시한 결과, 조지방, 등심 내의 수분, 육색, 적색도 그리고 황색도 등에서 유의적인 연관성을 관찰할 수 있었다. c.892A>G (Asp298Asn) 단일염기변이의 유전자형 빈도는 품종별로 차이가 났으며, A (Asn) 대립유전자의 빈도가 가장 높은 품종은 듀록으로 나타났고, G (Asp) 대립유전자의 빈도가 가장 높은 품종은 한국재래돼지로 조사되었다. c.892A>G (Asp 298Asn) 단일염기변이와 돼지 4 집단의 육질형질을 1,126두에서 분석한 결과, 등지방두께에 고도의 유의적인 효과를 관찰할 수 있었다(P<0.002). AA 유전자형을 가진 개체가 AG나 GG 유전자형을 가진 개체보다 등지방두께가 두꺼운 것을 확인할 수 있었다. 본 연구의 결과를 통해 MC4R 유전자 내의 c.892A>G (Asp298Asn) 단일염기변이는 돼지의 선발개량에 유전자표지인자로서 충분한 효과가 있음을 검증하였다.

오적산가미방(五積散加味方)이 고지방식이 유도 비만쥐의 지질대사, 항산화계 및 전염증성 cytokine 생산에 미치는 영향 (Effects of Ojeoksangamibang on the Lipid Metabolism, Anti-oxidation and Concentration of Proinflammatory Cytokines in Rat Fed High Fat Diet)

  • 공인표;박원형;차윤엽
    • 한방재활의학과학회지
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    • 제21권4호
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    • pp.23-40
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    • 2011
  • Objectives: This study was designed to examine the effects of extracts of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) on the lipid lowering, anti-oxidation and concentration of proinflammatory cytokines and was investigated on hyperlipidemic rats. Methods: Male rats weighing $182.39{\pm}4.71g$ were fed high fat diet for 8 weeks and 36 rats(above 400 g) were divided into 4 groups. Each of 9 rats was divided a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And we fed each experimental group of rats basal diet and administered an extract of Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) extracts(100 mg/kg, 200mg/kg, 300 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflmmatory cytokines, anti-oxidative activity and $TNF-{\alpha}$, Apo-B, Apo-E and leptin gene expression. Results: 1. Concentration of plasma free fatty(FFA) showed no significant difference in all the treatment groups. Concentration of plasma triglyceride(TG) showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 2. Concentration of plasma total cholesterol showed a significant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma low density lipoprotein(LDL)-cholesterol showed a Significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. Concentration of plasma high density lipoprotein(HDL)-cholesterol showed a significant increment in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group. 3. Concentration of liver total cholesterol showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. Concentration of liver TG showed a significant decrement in all Ojeoksangamibang groups than that of control group. 4. Concentration of plasma and liver thiobarbituric acid reactive substance(TBARS) showed a tendence to decrease in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups. 5. The values of glutathione peroxidase(GSH-Px), superoxide dismutase(SOD) and catalase(CAT) activity showed a significant increment in all Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups than that of control group. 6. The values of plasma aspartate aminotransferase(AST) and alanine aminotransferase(ALT) activity showed no significant different in all treatment group. 7. Concentration of plasma $interleukin(IL)-1{beta}$ showed no significant difference in all the treatment groups. Concentration of plasma IL-6 showed a significant decrement in the 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. Concentration of plasma tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ a siginifant decrement in the 200 and 300 mg/kg in Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) group than that of control group. However the concentration of plasma IL-10 in the 300 mg/kg Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant increment than that of control group. 9. In the analysis of reverse transcription-polymerase chain reaction(RT-PCR), gene expression of $TNF-{\alpha}$, Apo-B and Apo-E in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a lower expression than that of control group. However the gene expression of leptin showed no difference in the treatment groups. 10. The ratio of $TNF-{\alpha}$, Apo-B, and Apo-E per ${\beta}-actin$ expression in the Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) groups showed a significant decrement than that of control group. However The ratio of leptin expression per ${\beta}-actin$ expression showed no significant difference among all the treatment groups. Conclusions: According to above results, in lowering lipid effect, anti-oxidation and control of pro-inflammatory cytokines production, Ojeoksangamibang($W{\check{u}}j\bar{i}s\check{a}nji\bar{a}w\grave{e}if\bar{a}ng$) gives effect.