초록 ▼

Ⅳ. 연구개발결과
1. 24개의 능력 및 품성 관련 유전자들에 대한 생물정보학적 분석
2. 군견 자견적격심사를 통과한 개체군과 탈락한 개체군에서의 TH 유전자 내 VNTR변이양상 비교 분석
3. TH 유전자의 VNTR에 따른 서로 다른 allele의 서열 비교
4. 흰개미 탐지견 (삽살개) 표현형 50두에서 TH 유전자 분석
5. VNTR의 copy 수에 따른 인핸서 활성 비교
6. 다양한 개체에서의 품성 관련 유전자의 DMRs 분석(후성유전학적 연구)
7. MAOA 유전자의 견종별 발현양상 및

Ⅳ. 연구개발결과
1. 24개의 능력 및 품성 관련 유전자들에 대한 생물정보학적 분석
2. 군견 자견적격심사를 통과한 개체군과 탈락한 개체군에서의 TH 유전자 내 VNTR변이양상 비교 분석
3. TH 유전자의 VNTR에 따른 서로 다른 allele의 서열 비교
4. 흰개미 탐지견 (삽살개) 표현형 50두에서 TH 유전자 분석
5. VNTR의 copy 수에 따른 인핸서 활성 비교
6. 다양한 개체에서의 품성 관련 유전자의 DMRs 분석(후성유전학적 연구)
7. MAOA 유전자의 견종별 발현양상 및 CpG site 메틸화 양상 비교
8. HRM기법을 이용한 우수 특수목적견 특이적 메틸화 양상 분석
9. 서로 다른 두 견종의 근육 및 대뇌 조직에서 차등 발현하는 유전자의 정량적인 발현 및 프로모터 영역의 메틸레이션 양상 비교 · 분석
10. microsatelite 분석을 통해 우수 특수목적견 표현 형질 관련 DMRs 변이 지도 작성과 종합적 관계도 구축
11. 품성유전자 관련 개 특이적 miRNA 마커 및 기능 연구 (후성유전학적 연구)
12. 우수 특수목적견 특이적인 miRNA 발굴 및 발현양상 확인
13. 개 고관절탈구 관련 유전자 좌위에 대한 PCR-RFLP
14. 11개 단일염기변이 좌위에 대한 PCR-RFLP 검증 실험 및 유전자 분석 키트화
15. 13개 단일염기변이 좌위에 대한 PCR-RFLP 검증 실험 및 유전자 분석 키트화
16. 개 고관절이형성증 조기진단 10개 좌위 TAQMEN probe 키트화
17. 특수목적견의 품종 및 기능별 미토콘드리아 DNA 변이 확대 조사 (242두)
18. 개의 근육 조직 내에서 근지구력 및 피로회복과 연관된 차등 발현 유전자의 발굴 및 유전자 간의 네트워크 조사.
19. 개 미토콘드리아 DNA 내 조절영역(D-loop) 단일염기변이 확대 조사
20. 특수목적견의 임무능력 관련 유전자 마커 개발
21. 특수 목적견 (삽살개) 의 강건성 관련 유전자 마커 개발
22. 특수 목적견 섭식능력 연관 유전자 마커 발굴
23. 특수 목적견 체형(흉심, 체고, 체장) 연관 유전자 마커 발굴
24. 특수 목적견 청각인지능력 연관 유전자 마커 발굴
25. 특수 목적견 후각인지능력 연관 유전자 마커 발굴

Abstract ▼

Canine and human behavior are shaped by similar evolutionary processes, yet the identification of the behavioral phenotype is often difficult. A widely used method relies on breed stereotypes provided by experts such as dog trainers. To reveal a valid association between behavior and genetic factors

Canine and human behavior are shaped by similar evolutionary processes, yet the identification of the behavioral phenotype is often difficult. A widely used method relies on breed stereotypes provided by experts such as dog trainers. To reveal a valid association between behavior and genetic factors, an association study of behavioral phenotyping and genotyping is essential.
The variable number of tandem repeats (VNTR) in intron 4 of the tyrosine hydroxylase (TH) gene is related to two characteristics, namely, impulsive activity and inattention. The TH genes are defined by three genotypes: type 1/1 comprises a single copy of TH VNTR; type 1/2 is heterozygote for both short allele and long allele; and type 2/2 is the doubled form of TH VNTR, which leads to inattention and impulsive actions.The 1/1 and 1/2 genotype have been identified as biomarkers for concentration and impulsive action.
The military working dogs showed the 2/2 type of VNTR and heteroduplex. For the pass group, two dogs possessed 2/2 type (40 %), whereas three dogs were of the heteroduplex type (60 %). However, all members of fail group showed the 2/2 type (100%). These data indicate that repeat polymorphisms with behavioral phenotyping can identify military working dogs that would pass or fail the in-training examination.
Therefore, for the further study, we performed PCR amplification to detect the 1/2 genotypes in three canine breeds from Korea: Sapsaree (49), Poongsan (18), and Jindo (20). Our results indicate that the Sapsaree dog carries 27 % of the 2/2 genotype, 59 % of the 1/2 genotype, and 14 % of the 1/1 genotype; the Poongsan dog carries 11 % of the 2/2 genotype, 22 % of 1/2 genotype, and 67 % of 1/1 genotype; whereas the Jindo dog carries 10 % of the 2/2 genotype, 40 % of the 1/2 genotype, and 50 % of the 1/1 genotype. Taken together, our data indicate that the Jindo, Poongsan, and Sapsaree dogs harbor decreasing amounts of phenotypes responsible for inattention and impulsive behavior, in that order. Additionally, the Sapsaree genes harbor higher amounts of the 1/2 genotype than the Jindo and Poongsan genes, respectively.
Another study for working dogs are the genetic diversity and relationships among two groups of working dogs (pass and fail group in-training examination) were assessed based on 15 microsatellite markers in 25 individuals of working dogs (military watch dogs and Korean search dogs). For the 15 microsatellite markers, the values of allelic richness(AR)
ranged from 2.21 (pass group) to 1.60 (fail group) in military watch dogs, while AR ranged from 2.79 (pass group) to 2.72 (fail group) in Korean search dogs. Among 52 different alleles of military watch dogs, 22 alleles were detected in pass group only, while 8 alleles in fail group only. In case of Korean search dogs, 3 alleles were observed in pass group only, while 13 alleles in fail group only. These group-specific unique alleles reflect good biomarker for selecting working dogs (military watch dogs and Korean search dogs), indicating that those group specific microsatellite alleles could separate working dogs to be pass or fail group in outtraining dog population.
There are more than 400 pure dog breeds developed through intentional artificial selection and purebred breeding. Purebred animals have higher risk of inbreeding depression and hereditary diseases. We investigated the genetic diversity and structure of three dog
breeds in South Korea by using 12 microsatellite loci for one Korean native dog breed, Sapsaree, and two foreign breeds, German shepherd and Belgian Malinois. The mean allele number of nine loci across all dog breeds was 4.833, and the number of alleles per locus ranged from 2 to 8. The mean of expected and observed heterozygosity were 0.415 and 0.577, respectively. Sapsaree, Korean native dog, had higher level of genetic diversity than the foreign German shepherd and Belgian Malinois. The highest mean value of polymorphism information content was found in Sapsaree (0.480), followed by Belgian Malinois (0.373) and German shepherd (0.355). Pairwise genetic differentiation was estimated using fixation index FST. Sapsaree and German shepherd (FST = 0.2536) and Sapsaree and Belgian Malinois (FST = 0.2522) had very great genetic differentiation, while moderate level of genetic differentiation was observed between German shepherd and Belgian Malinois (FST = 0.1003). These genetic information and structure of the three dog breeds will be effective in conservation and preservation of the genetic diversity of the three dog breeds.
MicroRNAs (miRNAs) are known for their role in mRNA silencing via interference pathways. Repetitive elements (REs) share several characteristics with endogenous precursor miRNAs. In this study, 406 previously identified and 1,494 novel RE-derived miRNAs were sorted from the GENCODE v.19 database using the RepeatMasker program.They were divided into six major types, based on their genomic structure. More novel RE-derived miRNAs were confirmed than identified as RE-derived miRNAs. In conclusion, many miRNAs have not yet been identified, most of which are derived from REs.
The domestic dog species is divided into over 300 pure breeding populations known as breeds. Many breeds are characterized by reduced genetic diversity due to small founding population sizes, excessive use of popular sizes, and changes in breed popularity over time. Especially, most of working dogs has been exposed to diseases for genetic selection focused on specific skills. Human HD, referred to as developmental dysplasia of the hip, occurs with a frequency ranging from 5 to 13%. CHD is a observed in many breeds of dogs and in almost all large-sized breeds of dogs. The condition of CHD is remarkably similar in their clinical expression and pathogenesis in both man and dog. In Labrador retrievers used as guide dogs, CHD is regarded as a major disease. CHD is a genetic disease, but its diagnosis is based on a late onset clinical phenotype such as X-ray of hip joints. A genome-wide association study was performed using the Canine SNP20 BeadChip with the objective of identifying biomarkers to develop a concise and exact detection method for CHD at early ages. Forty eight LRs (26 males and 22 females) aged between 1-15 years old were assessed for BVA/KC scores measured by X-ray diagnosis . The SNP markers most associated with CHD group were found at 11 loci on chromosome 4 using the Canine SNP20 BeadChip.
Mitochondrial DNA (mtDNA) genome of the domestic dog (Canis familiaris) contains 16,727 bp with the Control Region (CR) spanning positions 15,458-16,727 (1,270 bp). Within mtDNA, CR, or D-loop, is generally one of the most divergent regions and is the most appropriate place to investigate variability. Samples from 242 domestic dogs of 8 different breeds were sequenced and subsequently examined for sequence variation in dog mtDNA. 8 dirrerent breeds are Sapsaree dog, German Shepherd dog, Belgium Malinois dog, Jindo Dog, Poongsan Dog, Labrador Retriever dog, DongGyeong dog, Beagle dog. Substantial variation was found both within and among breeds in the mtDNA derived from blood. The most variable sites observed were 15,639 in Sapsaree dog, 16,099 in German Shepher dog and Malinois dog, 15,650 in Jindo Dog, 15,653 in Poongsan Dog, 15,972 in Labrador Retriever dog, 16,037 in DongGyeong dog, and 15,643, 15,650, 15,664, 15,816, 15,972, and 16,028 in Beagle dog. These were consistent with other reports on variable positions in the dog mtDNA genome.
DNA microarrays can give global transcriptional views of cellular responses to disease, development, nutrition, and other biological states. They can be used to elucidate biological networks, develop diagnostics, and identify genetic targets and molecular mechanisms. The technology is widely used and can be a useful complement to‘‘disease-centric’’ focused arrays. Array-based transcript profiling experiments enable the detection of thousands of gene expression patterns in a single experiment. Using the microarray technology, the objective of this study was to identify genes and functional expressed in muscles tissue in dogs. We found 18 expressed genes in canine muscle tissue.These 18 expressed genes was associated with calcium signal pathway, cell communication, ECM-receptor interaction, fatty acid metabolism, glutathione metabolism, glycerolipid metabolism, insulin signaling pathway and neurodegenerative disorders. The calcium signal pathway was associated with TACR2 gene. The cell communication and ECM-receptor interaction was associated with VTN gene. The fatty acid metabolism and glycerolipid metabolism was associated with ADH4 gene. The glutathione metabolism was associated with GSTM3 gene. The insulin signaling pathway was associated with CRKL and PPARGC1A gene. The neurodegenerative disorders was associated with APOE gene. These results represented that genes and muscle may be important relation.
Genetic characteristics of height and weight through analysis of high-density SNP 170K chip were significantly associated with SNPs of 26 loci. These SNPs of 14 loci for height of dogs were located on chromosomes 20 and 34, and SNPs of 12 loci for weight of dogs were located on chromosomes 3, 15 and 35.