Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Detection of a Large White-Specific Duplication in D-loop Region of the Porcine MtDNA

돼지 mtDNA D-loop 지역의 Large White 특이 중복현상 탐지

  • Kim, Jae-Hwan (National Institute of Animal Science, R. D. A.) ;
  • Han, Sang-Hyun (National Institute of Animal Science, R. D. A.) ;
  • Lee, Sung-Soo (National Institute of Animal Science, R. D. A.) ;
  • Ko, Moon-Suk (National Institute of Animal Science, R. D. A.) ;
  • Lee, Jung-Gyu (Division of Applied Life Science, Gyeongsang National University) ;
  • Jeon, Jin-Tae (Division of Applied Life Science, Gyeongsang National University) ;
  • Cho, In-Cheol (National Institute of Animal Science, R. D. A.)
  • 김재환 (농촌진흥청 국립축산과학원) ;
  • 한상현 (농촌진흥청 국립축산과학원) ;
  • 이성수 (농촌진흥청 국립축산과학원) ;
  • 고문석 (농촌진흥청 국립축산과학원) ;
  • 이정규 (경상대학교 응용생명과학부) ;
  • 전진태 (경상대학교 응용생명과학부) ;
  • 조인철 (농촌진흥청 국립축산과학원)
  • Published : 2009.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

The entire D-loop region of the porcine mitochondrial DNA (mtDNA) was amplified from six pig breeds (Landrace, Duroc, Large White, Korean native pig, Berkshire, and Hampshire) using a primer set designed on the basis of reported porcine mtDNA sequences. From analyses through cloning, DNA sequencing and multiple sequence alignment, an 11-bp (TAAAACACTTA) duplication was observed after known tandem repetition in the D-loop region, which promoted hetroplasmy in mtDNA. Although the existence of the 11-bp duplication has been previously reported in Duroc and Japanese native pigs, there have not been any attempts to know the characteristics of this duplication in other breeds so far. A 150 bp fragment containing the 11-duplication was amplified and typed by polyacrylamide gel electrophoresis (PAGE). All Large Whites had two duplication units and Duroc showed heteromorphic patterns, 11.2% (9/80) of the animals had the 11-bp duplication in total. On the other hand, Landrace, Berkshire, Hampshire and Korean native pigs were non-duplicated. This result showed that the 11-bp duplication could be used as a breed-specific DNA marker for distinguishing pure Landrace and Large White breeds.

돼지 6품종(Landrace, Duroc, Large White, 한국재래돼지, Berkshire, Hampshire)을 대상으로 기존에 보고된 서열을 바탕으로 제작한 primer를 이용하여 mtDNA D-loop 전체영역을 증폭하였다. 증폭된 PCR product를 cloning 및 DNA sequencing, 다중염기서열비교를 통하여 분석한 결과, mtDNA에서 heteroplasmy가 나타나는 D-loop 내 tandem repeat region 이후에 11-bp 중복이 존재하는 것을 확인하였다. 이런 중복현상은 일본재래돼지와 Duroc에서 보고되었지만, 이를 이용한 돼지 품종별 중복현상의 빈도 및 분포에 관한 연구는 이루어져있지 않다. 품종별 11-bp 중복현상을 분석하기 위해서 6품종을 대상으로 중복지역을 포함한 약 150 bp 절편을 증폭하였으며, PAGE 방법을 통하여 분석하였다. 그 결과 본 연구에서 사용한 품종들 중 모든 Large White에서 중복현상이 발생하는 것을 확인하였으며, Duroc인 경우 11.2% (9/80)에서 중복현상이 확인되었다. 반면에 Landrace, 한국재래돼지, Berkshire 및 Hampshire에서는 전혀 발견되지 않았다. 이런 결과로서, 11-bp 중복현상의 분석은 현재 구별이 불가능한 Landrace와 Large White를 구별할 수 있는 유용한 DNA marker로서 사용이 가능할 것이다.

Keywords

References

  1. Avise, J. C. 1986. Mitochondrial DNA and evolutionary genetics of higher animals. Philos. Trans. R. Soc. Lond. B 312, 325-342 https://doi.org/10.1098/rstb.1986.0011
  2. Birren, B., E. D. Green, S, Klapholz, R. M. Myers, and J. Roskams. 1997. Genome analysis: A laboratory manual. Cold Spring Harbor Laboratory Press, USA
  3. Cann, R. L., W. M. Brown, and A. C. Wilson. 1984. Polymorphic sites and the mechanism of evolution in human mitochondrial DNA. Genetics 325, 31-36
  4. Cho, I. C., Y. H. Jung, J. K. Jung, P. N. Seong, B. W. Kim, J. G. Lee, and J. T. Jeon. 2003. Genetic Variation of Mitochondrial DNA in Duroc (Sus Scrofa) Using Single Stranded Conformation Polymorphism Analysis. J. Anim. Sci. & Technol. (Kor.) 45, 911-916 https://doi.org/10.5187/JAST.2003.45.6.911
  5. Cook, C. E., Y. Wang, and G. Sensabaugh. 1999. A mitochondrial control region and cytochrome b phylogeny of Sika Deer (Cervus Nippon) and report of tandem repeats in the control region. Mol. Phylogenet. Evol. 12, 47-56 https://doi.org/10.1006/mpev.1998.0593
  6. Ghivizzani, S. C., S. L. D. Mackay, C. S. Madsen, P. J. Laipis, and W. W. Hauswirth. 1993. Transcribed heteroplasmic repeated sequences in the porcine mitochondrial DNA D-loop region. J. Mol. Evol. 37, 36-47
  7. Giles, R. E., H. Blanc, H. M. Cann, and D. C. Wallace. 1980. Maternal inheritance of human mitohondrial DNA. Proc. Natl. Acad. Sci. 77, 6715-6719 https://doi.org/10.1073/pnas.77.11.6715
  8. Jiang, S. W., E. Giuffra, L. Andersson, and Y. Z. Xiong. 2001. Molecular Phylogenetics relationship between six Chinese native pig breeds and three Swedish pig breeds from mitochondrial DNA. Yi. Chuan. Xue. Bao. 28, 1120-1128
  9. Kijas, J. M. H., M. Moller, G. Plastow, and L. Andersson. 2001. A frameshift mutation in MC1R and a high frequency of somatic reversions cause black spotting in pigs. Genetics 158, 779-785
  10. Kijas, J., R. Wales, A. Trnsten, P. Chardon, M. Moller, and L. Andersson. 1998. Melanocortin receptor 1 (MC1R) mutations and coat color in pig. Genetics 150, 1177-1185
  11. Kim, K. I., J. H. Lee, K. Li, Y. P. Zhang, S. S. Lee, J. Gongora, and C. Moran. 2002. Phylogenetic relationships of Asian and European pig breeds determined by mitochodrial DNA D-loop sequence polymorphism. Anim. Genet. 33, 19-25 https://doi.org/10.1046/j.1365-2052.2002.00784.x
  12. Lin, C. S., Y. L. Sun, and C. Y. Liu. 1999. Complete nucleotide sequence of pig (Sus scrofa) mitochondrial genome and dating evolutionary divergence with Artiodactyla. Gene 236, 107-114 https://doi.org/10.1016/S0378-1119(99)00247-4
  13. Marklund, S., J. Kijas, H. Rodriguez-Martinez, L. Rnnstrand, K. Funa, M. Moller, D. Lange, I. Edfors-Lilja, and L. Andersson. 1998. Molecular basis for the dominant white phenotype in the domestic pig. Genome Res. 8, 826-833
  14. Mergen, H., R. Oner, and C. Oner. 2004. Mitochondrial DNA sequence variation in the Anatolian Peninsula (Turkey). J. Genet. 83, 39-47 https://doi.org/10.1007/BF02715828
  15. Miracle, A. L. and D. E. Campton. 1995. Tandem repeat sequence variation and length heteroplasmy in the mitochondrial DNA D-loop of the threatened Gulf of Mexico sturgeon, Acipenser oxyrhynchus desotoi. J. Hered. 86, 22-27
  16. Okumura, N., N. Ishiguro, M. Nakano, H. Katsuya, A. Matsui, and M. Sahara. 1996. Geographic population structure and sequence divergence in the mitochondrial DNA control region of the Japanese wild boar (Sus scrofa leucomystax), with reference to those of domestic pigs. Biochem. Genetics 34, 179-189 https://doi.org/10.1007/BF02407018
  17. Thompson, J. D., T. J. Gibson, F. Plewniak, F. Jeanmougin, and D. G. Higgins. 1997. The ClustalX windows interface: flexible strangies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25, 4876-4882 https://doi.org/10.1093/nar/25.24.4876
  18. Wilson, A. C., L. Cann, S. M. Carr, M. George, and U. B. Gyllensten. 1985. Mitochondrial DNA and two perspectives on evolutionary genetics. Biol. J. Linn. Soc. 26, 375-400 https://doi.org/10.1111/j.1095-8312.1985.tb02048.x
  19. Xu, X. and U. Arnason. 1994. The complete mitochondrial DNA sequence of the horse, Equus caballus: extensive heteroplasmy of the control region. Gene 148, 357-362 https://doi.org/10.1016/0378-1119(94)90713-7
  20. Yu, L., Q. W. Li, O. A. Ryder, and Y. P. Zhang. 2004. Phylogeny of the bears (Ursidae) based on nuclear and mitochodrial genes. Mol. Phylogenet. Evol. 32, 480-494 https://doi.org/10.1016/j.ympev.2004.02.015