초록 ▼

○ 연구결과
1. 제주마 유전적 특성 파악
- RAPD PCR을 이용한 마 품종 유전특성 확립 총 22개 priner
- 제주마(JNH)는 더러브렛종과 다른 고유한 유전적 특성이 있는 것으로 확인
- SCAR primer을 활용한 제주마 특이 인자 식별 가능
2. 제주마의 친자감별 및 개체식별 기법 확립
- MS 좌위를 통한 개체식별기법의 개발과 실용화가 가능케됨.
- 유전현상을 보이면서 다형현상이 높은 10개의 MS좌위에 대한 획득성과는 제주마에 대한 분자유전학적인 친자판정을 가능케 함.

○ 연구결과
1. 제주마 유전적 특성 파악
- RAPD PCR을 이용한 마 품종 유전특성 확립 총 22개 priner
- 제주마(JNH)는 더러브렛종과 다른 고유한 유전적 특성이 있는 것으로 확인
- SCAR primer을 활용한 제주마 특이 인자 식별 가능
2. 제주마의 친자감별 및 개체식별 기법 확립
- MS 좌위를 통한 개체식별기법의 개발과 실용화가 가능케됨.
- 유전현상을 보이면서 다형현상이 높은 10개의 MS좌위에 대한 획득성과는 제주마에 대한 분자유전학적인 친자판정을 가능케 함.
- 제주마의 혈통확립을 위한 친자판정의 체계를 완성하였다. 이로서 친자판정에 따른 시비와 혈통추적의 문제가 해결됨.
- 본 연구결과물과 분석방법에 대한 기법은 제주도 축산진흥원에서 진행되고 있는 제주마 등록관리사업의 효율성에 가장 기본적이면서도 필수적인 수단을 제공하게 됨.
3. 제주마의 혈통등록 체계 확립
- 제주마 일반 외모심사 기준 완료
- 제주마 연령별 발육성적 조사 완료
- 제주마 혈통등록 규정 확립
- 기초 및 혈통등록 266두 완료

Abstract ▼

Ⅳ Results and Suggestion on Utilization of Results
1. Idetification of genetic markers in Horse
RAPD was performed to estimate the genetic characteristics between breeds and to obtain the specific DNA marker for Jeju native horse and other breeds. of the initial 1980 random primers, 1,539 prim

Ⅳ Results and Suggestion on Utilization of Results
1. Idetification of genetic markers in Horse
RAPD was performed to estimate the genetic characteristics between breeds and to obtain the specific DNA marker for Jeju native horse and other breeds. of the initial 1980 random primers, 1,539 primers produced clear and intense bands following PCR amplification. Despite the large number of primers screened, most of the primers generating interpretable amplification profiles showed monomorphic pattern among horse breeds. Amomg 1539 primers 574 primers were selected to identify the breed specific DNA marker and for the genetic difference between breeds and whithin. a total of 7,857 bands were generated by 574 primers and the number of amplified produfcts per each primer ranged from 3 to 14 with an average of 5.1. The size of the amplified fragments represented from 150bp to 2600bp.
Genetic similarities was calculated with 13 microsatellite Allele frequencies in comparison of six horse breeds. and were used to estimate expected heterozyosity(He), genetic distance (Ds). The expected heterozyosity(He) 13 microsatekkite loci for each breeds were 0.700 for JNH, 0.728 for Tho, 0.797 for Mong, 0.794 for JRH, 0.39 for Jap and 0.705 for Qua. This matrix was used to generate dendrogram by UPGMA methods.
The genetic similarity coefficient between Jeju native horse and chinese was 0.175, 0.782 for Jeju native horse and thoughbred, 0.666 for thoroughbred and japan horse, 0.413 for chinese and japan horse, 0.782 and was the highest value in comparison of pair breeds.
Comparing the pooled DNA from Jeju Native Horse and Thoroughbred, we found 25 primers which identified markers present in the pooled DNA from breed but absent in the other breed. Among 290 random primers, 157 primers were Thoroughbred specific and 133 primers were Jeju Native Horse specific. Testing individual horse revealed that 5 marker showed the similar band pattern between Jeju Native Horse and Thoroughbred. However, 22 marker were wholly absent in breed while present in the
other breed( MG0244, MG0162, MG0003, MG0065, MG0034, MG0055, MG5086, MG5153.
MG0085, MG0326, MG6085, MG0446, MG0177, MG0562, MG0171, MG0139, MG0162, MG0443, MG02043, MG0030, MG0126 MG0081.
We cloned and sequenced for specific RAPD fragment for 24 clones. These results demonstrates that non repetitial sequence. Therefore, these RAPD bands specifie to breed have potential possibility for genetic marker for specific breed. PAPD bands specifie to breed have potential possibility for genetic marker for specific breed. RAPD bands specific for could be used to distinguish Jeju Native Horsethe from Thoroughbred or other crossbred.
we concluded that RAPD screening of DNA pools of six populaton was an effective approach for identifying markers which distinguish each horse breeds. This approach could be converted to the system based on sequence characterized amplified region (SCAR) and would avoid the technical difficulties inherent in RAPD testing. The power of the test would depend upon the number of markers used and of the relative frequency of the markers within each populations being compared. Increasing the number of markers would make the system even more powerful and this would be particularly important for idetification of JNH.
2. Development of Microsatellite DNA markers for
Individual Identification and Parentage Verification in CheJu Native Horse
After the candidate microsatellite markers were analysed and evaluated about the Mendelian inheritance and polymorphisms, a total of ten microsatellite markers(ASB25, HMS5, HTG7, UCDEQ411, UM010, UM012, VHL20, VHL209, VIAS-H39 and VIAS-H64) were selected and determined in this research. Total number of alleles over 10 microsatelliet loci was 68(mean number of alleles per locus was 6.8).
To test the usefulness about the set of selected microsatellite loci in individual identification system, microsatellite genotyes of 100 horses which were gathered from candidate horses for registration were compared each other individual genotype. Any pairs of individuals did not have the same genotypes, indicating that the set of selected microsatellite loci has a good power of individual identification.
A total of 40 foals and 44 individuals for candidate parents were typed for their genotypes in each microsatellite locus and demonstrated to test the usefulness of this set of microsatelliet markers in parentage verification. Given two parents and one offspring the probabilities of exclusion were 1.00000 in the horse group of farms and 0.99996 in the group of Jeju Institute. Using the allele system of microsatellite loci which are developed in these studies allows individuals to be a quick and clear inclusion, as well as exclusion in parentage testing. An individual can be identified positively as the parent rather than merely being eliminated as a positively without critical errors.
The developed system of parent verification will be used to genetically identify individuals and verify parentage of Cheju horses under the registration system of Jeju Institute.
3. Development of Pedigree System of Jeju Native Horse
The general standard of the external shape was established by the documentary records. It was able to use for the Cheju horse registration.

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 요 약 문 ... 3
• SUMMARY ... 7
• CONTENTS ... 12
• 목 차 ... 13
• 제 1 장 연구개발과제의 개요 ... 14
• 제 1 절 연구개발의 필요성 ... 14
• 가. 기술적측면 ... 14
• 나. 경제‧산업적 측면 ... 15
• 다. 사회․문화적 측면 ... 15
• 제 2 절 연구개발의 목표와 범위 ... 16
• 제 2 장 국내외 기술개발 현황 ... 17
• 1. DNA 다형현상을 이용한 분자육종 기술개발 연구 ... 17
• 제 3 장 연구개발수행 내용 및 결과 ... 18
• 제1절 : 제주마의 유전특성구명 및 유전표지인자 개발 ... 18
• 1. 추진 방법 ... 18
• 2. 결과 및 고찰 ... 19
• 제 2절: 제주마의 친자감별 및 개체식별 기법확립 ... 88
• 1. 추진 방법 ... 88
• 2. 결과 및 고찰 ... 92
• 제 3 절 : 제주마의 혈통등록 체계확립 ... 102
• 1. 추진 전략 및 결과 ... 102
• 제 4장 목표달성도 및 관련분야에의 기여도 ... 106
• 1. 연차별 연구개발목표와 내용 ... 106
• 2. 연구개발목표의 달성도 ... 108
• 3 관련분야의 기술발전에의 기여도 ... 109
• 제 5장 연구개발결과의 활용계획 ... 109
• 6장 참고 문헌 ... 110
• 끝페이지 ... 111