Journal of Animal Science and Technology

  • 제49권5호
  • /
  • Pages.559-568
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  • 2007
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  • 2672-0191(pISSN)
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  • 2055-0391(eISSN)
한국축산학회 (Korean Society of Animal Sciences and Technology)
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Quantitative Oligonucleotide Ligation Assay(qOLA)를 이용한 Landrace 품종의 KIT 유전자 반복수 변이 탐지

Detection of Copy Number Variation of the KIT Gene in the Landrace Breed using an Quantitative Oligonucleotide Ligation Assay(qOLA)

  • 서보영 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 김재환 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 남덕우 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 유채경 (경상대학교 농생명학부) ;
  • 이상호 (경남동물과학기술) ;
  • 이재봉 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 임현태 (경상대학교 대학원 응용생명과학부(BK21)) ;
  • 정은지 (경상대학교 농생명학부) ;
  • 조인철 (농촌진흥청 난지농업연구소) ;
  • 허강녕 (축산과학원 동물유전체과) ;
  • 전진태 (경상대학교 대학원 응용생명과학부(BK21))
  • Seo, B.Y. (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Kim, J.H. (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Nahm, D.W. (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Yoo, C.K. (Division of Agriculture and Life, Gyeongsang National University) ;
  • Lee, S.H. (Gyeongnam Animal Science Technology) ;
  • Lee, J.B. (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Lim, H.T. (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University) ;
  • Jung, E.J. (Division of Agriculture and Life, Gyeongsang National University) ;
  • Cho, I.C. (National Institute of Subtropical Agriculture, R. D. A.) ;
  • Heo, K.N. (National Institute of Animal Science, R. D. A.) ;
  • Jeon, J.T. (Division of Applied Life Science(BK21), Graduate School of Gyeongsang National University)
  • 발행 : 2007.10.31
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초록

최근 들어 유전자 또는 DNA 분절의 반복수 변이 (CNV)에 관한 연구가 다수 수행되었으며, 그 분석 방법 및 기기 또한 다양하게 발달되었다. 본 연구는 Landrace 품종의 KIT 유전자 CNV 탐지를 위하여 다른 분석 방법들에 비하여 정확도가 높은 정량적 OLA 기법(qOLA)을 이용하였다. qOLA와 pyrosequencing assay의 조합하여 분석한 결과를 Landrace 44두에 대한 좌표 분석을 한 결과 I1/I1 또는 I3/i(IBe), I1/I2 또는 I3/IP, I1/I3, I1/IP 또는 I2/i(IBe), I2/I2, I2/IP의 6 genotype으로 분류되었으며, PROC FASTCLUS procedure을 이용한 통계 분석과 좌표 분석을 상호 비교한 결과 genotype의 분류가 100% 일치하였다. 기기간의 차이점을 조사하기 위해 qOLA_3100과 qOLA _3130의 관측치 비교를 실시한 결과 동일한 genotype 분류결과를 얻었다. 또한 정밀성 및 정확도 비교에서 qOLA_3100의 경우 표준편차와 변이계수의 평균이 2.33과 4.10로 qOLA_3130(2.67과 4.81)에 비하여 낮게 나타났다. qOLA 반응전 PCR 산물에 대하여 proteinase K 처리효과를 분석한 결과 pro- teinase K를 처리한 경우 전기영동 분석시 noise peak들이 제거되었으며 각 genotype의 이론적 비율에 보다 정확히 일치하였다.

Recently, copy number variations (CNV) of genes or genomic segments have been intensively studied and various analysis methods have been developed. In this study, quantitative oligonucleotide ligation assay (qOLA) was applied to investigate CNV of KIT gene in the Landrace breed. A combined assay using qOLA and pyrosequencing, 6 genotype classes, I1/I1 or I3/i (IBe), I1/I2 or I3/IP, I1/I3, I1/IP or I2/i (IBe), I2/I2and I2/IP, were identified from 44 Landrace pigs. Genotype assignment using grouping features of measurements on a scatter plot showed 100% agreement with those using a statistical assignment by PROC FASTCLUS procedure implemented in the SAS package. Two versions (3100 and 3130) of ABI sequencers gave the same genotyping results, indicating there was no influence on qOLA by different versions of instrument, however, the means of standard deviation and coefficient of variation from the qOLA on a ABI 3130 (2.33 and 4.10) was lower than those from the qOLA on a ABI 3100 (2.67 and 4.81). Effect of proteinase K treatment on the PCR product followed by qOLA was very clear because noise peaks were disappeared and the observed ration fit better to the reference ratio corresponding to each genotype.

키워드

참고문헌

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