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Genetic Variation of Mitochondrial DNA in Duroc (Sus Scrofa) Using Single Stranded Conformation Polymorphism Analysis

Single Stranded Conformation Polymorphism 분석에 의한 돼지 Duroc 품종의 미토콘드리아 DNA 유전적 변이

  • Cho, I.C. (National Jeju Agricultural Experiment Station, R.D.A.) ;
  • Jung, Y.H. (National Jeju Agricultural Experiment Station, R.D.A.) ;
  • Jung, J.K. (National Jeju Agricultural Experiment Station, R.D.A.) ;
  • Seong, P.N. (National Jeju Agricultural Experiment Station, R.D.A.) ;
  • Kim, B.W. (Division of Applied Life Science, Gyeongsang National University) ;
  • Lee, J.G. (Division of Applied Life Science, Gyeongsang National University) ;
  • Jeon, J.T. (Division of Applied Life Science, Gyeongsang National University)
  • 조인철 (농촌진흥청 제주농업시험장) ;
  • 정용환 (농촌진흥청 제주농업시험장) ;
  • 정진관 (농촌진흥청 제주농업시험장) ;
  • 성필남 (농촌진흥청 제주농업시험장) ;
  • 김병우 (경상대학교 응용생명과학부) ;
  • 이정규 (경상대학교 응용생명과학부) ;
  • 전진태 (경상대학교 응용생명과학부)
  • Published : 2003.12.31

Abstract

The mitochondrial DNA(mtDNA) D-loop region was amplified from Duroc(Sus scrofa) by polymerase chain reaction(PCR). The oligonucleotide primer used to amplify the Sus scrofa mtDNA D-loop region was designed using tRNA-Pro and tRNA-Phe sequence in mtDNA regions highly conserved in many other animal species. There were 1,145 base pairs(bp) in the D-loop region. The middle of the region contained 10 tandem repeat of an 10-bp Sus scrofa-specific sequence, TACACGTGCG. We designed primers for PCR-mediated single stranded conformation polymorphism(SSCP) analysis that amplified a 345 bp fragment, which contained the most variable region according to our sequencing data. SSCP analysis of denatured amplification products was carried out by polyacrylamide(8%) gel electrophoresis followed by ethidium bromide staining. The SSCP analysis identified two band patterns(A and B) and comparision of these two nucleotide sequences identified 21 base substitutions. These results show that SSCP analysis of the D-loop region is useful for detecting the genetic polymorphism.

Keywords

D-loop;SSCP;Sus scrofa;Tandem repeat

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