Browse > Article
http://dx.doi.org/10.5352/JLS.2005.15.3.474

Development of Sequence Characterized Amplified Regions (SCAR) Showing for Cheju Native Horse  

Cho Byung Wook (Department of Animal Science, Miryang National University)
Publication Information
Journal of Life Science / v.15, no.3, 2005 , pp. 474-478 More about this Journal
Abstract
This study was conducted to analyze genetic characteristics and to develop the specific marker for Cheju native horse (Coo) at the level of sequence characterized amplified regions (SCARs). We collected blood samples from Cheju native horse and Thoroughbred horse (Th) and obtained genomic DNA from the blood of 50 individuals randomly selected within the breeds. Seven hundred primers were chosen randomly and were used to examin the polymorphism and 40 kinds of primers showed polymorphic RAPD band patterns between two breeds. Thirty primers of them showed horse specific bands. With the primer MG 30, amplified band of 2.0 kb showed the specificity to Cheju native horse (Cnh). Additionally MG 53 detected the thoroughbred horse (Th) specific markers at size of 2.3 kb. As the next, 2.3 kb band from MG 53 was checked with the all individuals from all the breeds of this study, and it maintained the reproducible breed specificity to thoroughbred horse (Th). With this results, 2.3 kb band was cloned into plasmid vector and sequenced bidirectionally from both ends of the cloned fragment. With the obtained sequences 10 nucleotide extended primers including the original arbitray primer were designed as a SCARs primer. Finally, the primer with extended sequence showed the reproducible breed differentiation pattern and it was possible to identify Cheju native horse (Cnh) from other breeds. The SCARs marker 2.3 kb from MG 53 could be used to identify Cheju native horse (Cnh) for not only registration but also horse breeding programe.
Keywords
Cheju native horse (Cnh); DNA Marker; Thoroughbred (Th); RAPD-PCR; SCAR;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Bailey, E. and T. L. Lear. 1994. Comparison of Thoroughbred and Arabian horse using RAPD markers. Anim Genet. 25, 105-108   DOI   ScienceOn
2 Kim, K. I., Y. H. Yang, S. S. Lee, C. Park, R. Ma, J. J. Bouzat and H. A. Lewin. 1999. Phylogenetic relationships of Cheju horses to other horse breeds as determined by mtDNA D-loop sequence polymorphism. Anim. Genet. 30, 102-108   DOI   ScienceOn
3 Micheli, M. R., R. Bova., E. Pascale and E. D' Ambrosio. 1994. Reproducibility of Random Amplified Polymorphic DNA (RAPD) analysis among laboratories. PCR Method. Appl. 2, 341-345
4 Myers, R. M., T. Maniatis and J. S. Lerman. 1987. Detection and localization of single base changes by denaturing gradient gel electrophoresis. Methods Enzymol. 155, 501-527   DOI
5 Paran, I. and R. Michelmore, 1993. Development of reliable PCR-based markers linked to downy mildew resistance genes in lettuce. Theor. Appl. Genet. 85, 985-993
6 Plotsky, Y., M. G. Kaiser and S. J. Lamonts. 1995. Genetic characterization of highly inbred chicken lines by two DNA methods: DNA fingerprinting and polymerase chain reaction using arbitrary primers. Anim. Genet. 26, 163-170   DOI   ScienceOn
7 Sambrook, J., E. F. Fritsch and T. Maniatis. 1989. Molecular cloning of a laboratory manual. 2nd Ed. Cold Spring Habor Laboratory
8 Williams, J. G. K., A. R Kubelik, J. A. Rafalski and S. V. Tingy. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18, 6531-6535   DOI   ScienceOn
9 Waugh, R. and W. Powell. 1992 using RAPD markers for crop improvement. TIBTECH 10, 186-191   DOI   ScienceOn
10 Ishida, N., T. Hasegawa, T. Oyunsuren and H. Mukoyama. 1996. PCR-RFLP analysis of the cytochrome b gene in horse mitochondrial DNA. Anim. Genet. 27, 359-363   DOI   ScienceOn
11 Kemp. S. J. and A. J. Teale. 1994. Randomly primed PCR amplification of pooled DNA reveal polymorphism in a ruminant repetitive DNA sequence which differentiated Bas indicus and Bas taurus, the major subspecies of domestic cattle. Anim Genet. 25, 82-88
12 Hayash, K. 1991. PCR_SSCP: A simple and sensitive method for detection of mutations in genomic DNA. PCR Meth. Appl. 1, 34-38   DOI
13 Cargill, S. L., G. B. Anderson and J. F. Medrano. 1995. Development of a species-specific marker using RAPD analysis to distinguish between sheep and goats. Anim. Biotechnol. 6, 93-100   DOI   ScienceOn
14 Cho, B. W., K. C. Hwang, H. K. Lee, K. J. Lee and J. Y. Han. 1997. Application of DNA markers to characterize cattle breed using random amplified polymorphic DNAs. Anim. Genet. and Breeding. 1, 39-48
15 Hong, Y. H., J. C. Cheong, T. H. Kim, H. B. Kim, D. H. Yoon, H. S. Kim, B. W. Cho and J. Y. Han. 1998. Development of sequence characterized amplified regions (SCARs) showing Korean cattle (Hanwoo) specificity. Anim. Genet. and Breeding. 2, 107-114
16 Jeffrey, A. J., V. Wilson and S. J. Thein. 1985. Individual specific fingerprints of human DNA. Nature 316, 76   DOI   ScienceOn
17 Welsh, J. and M. McClelland. 1990. Fingerprinting genomes using PCR with arbitary primers. Nucleic Acids Res. 18, 7213-7218   DOI   ScienceOn