Abstract
This experiment was carried out to analyze genetic characteristics and to develop the breed specific DNA marker for Cheju-native horse. If this marker contains high repetitive sequences, it is possible to convert a RAPD marker of interest into a single-locus PCR marker called a sequence characterized amplified region(SCAR). Twenty six Cheju-native horse and Fifty thoroughbred genomic DNA were pooled and PCR. were accomplished using 800 random primers. Comparing the pooled DNA from Cheju-native horse and thoroughbred, we found 9 primers which identified markers present in the pooled DNA from breed but absent in the other breed. Among 9 random primers, 6 primers were thoroughbred specific and 3 primers were Cheju-native horse specific. Testing individual horse revealed that 5 marker showed the similar band pattern between Cheju-native horse and Thoroughbred. However, 4 marker were wholly absent in breed while present in the other breed. UBC $126_{3500bp}$, UBC $162_{500bp}$, and UBC $244_{1200bp}$ was detected only Thoroughbred and UBC $562_{560bp}$was detected Cheju-native horse, respectively. After determining of the cloned breed-specific fragment sequence, we designed the SCAR-primers and carried out PCR. Compared to random primer, RAPD-SCAR primer didn't show significantly higher specific band. However, RAPD analysis is useful for genetic characterization of Cheju-native horse.
본 연구는 short oligonucleotide primer를 이용하여 마 품종간 유전 분석을 실시 하고자 PCR증폭 기법을 확립하고, 확립된 기술을 이용하여 제주도에 사육중인 천념기념물 347호로 등록된 제주말과 경주마로 잘 알려진 더러브렛간의 유전적인 다양성을 분석한 결과 마 품종간 차이를 보이는 DNA marker는 9개의 primer에서 확인되었으며, 이중 6개의 primer에서 더러브렛 특이 밴드와 나머지 3개에서 제주 마 특이 RAPD 밴드가 확인되어 cloning과 sequencing후에 SCAR primer를 제작하여 마 품종 식별에 활용할 수 있을 것으로 사료되며, 본 연구결과 RAPD표지인자는 마 품종간의 유전 분석에 매우 유용한 것으로 판단되었다.