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

국문 초록 (Abstract)

본 연구는 RAPD 기법을 이용한 종 특이 marker 개발 및 이 marker의 SCAR marker로의 개발을 목표로 수행되었다. Random primer 700개에 대하여 PCR 수행결과, 품종간, 개체간에 많은 다형성이 관찰되었으며 품종특이적인 양상을 나타내는 MG30, MG53의 primer는 각각 2.0kb, 2.3kb의 위치에서 제주말과 더러브렛종의 특이적인 RAPD 단편을 나타내었다. 이들 단편들 중 품종 특이적인 단편을 클로닝한 후 random primer가 포함된 부분의 염기서 열을 결정하였다. 10 bp의 RAPD random primer에 10bp의 염기를 추가하여 SCAR primer를 제작하였다. SCAR marker의 수행결과 RAPD marker와 같은 2.3kb, 2.0kb의 크기에서 제주마와 더러브렛종에 특이적인 하나의 밴드가 증폭되었다. 따라서 이 Cnh-SCAR marker는 보다 안정적이고 재현성 있는 marker로서 사용이 가능하여 제주말의 판별에 유용하게 사용될 수 있을 것이다.

다국어 초록 (Multilingual 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.

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