초록

Ⅳ. 연구개발결과
○ 한국재래돼지 지방형질 연관 유전자 물리지도 및 유전자 변이 발굴
○ 한국 재래견 경제형질관련 유전자지도 작성 및 변이 발굴
○ 한국재래가축 유전체 정보의 통합검색 활용체계 구축

Abstract ▼

The pig, Sus scrofa, was domesticated from wild boar subspecies in Asia and Europe. It was suggested that the domestication of pig was taken at multiple centers across Eurasia by analysis of mitochondria DNA sequences. The pig is one of livestock for meat resource in the world and is considered as a

The pig, Sus scrofa, was domesticated from wild boar subspecies in Asia and Europe. It was suggested that the domestication of pig was taken at multiple centers across Eurasia by analysis of mitochondria DNA sequences. The pig is one of livestock for meat resource in the world and is considered as an animal model for human health. Fatness trait of the pig is one of the crucial parameters determining meat quality of pork and is an important point as an animal model of obesity in humans. In decade, many efforts have been made to identify the effective chromosomal regions for meat quality such as backfat thickness (BFT) and intramuscular fat content (IMF) by quantitative trait loci (QTL) analysis on pig chromosome 6 (SSC6). Recent QTL analyses revealed that the region between S0228 and SW1881 might be highly significant QTL affecting fatness and meat quality traits on SSC6. The genomic region between S0228 and SW1881 corresponds to human chromosome 1 (65 Mb 85 Mb) and 18 (37 Mb 38 Mb) including breakpoint region between human chromosome 1 and 18. In this study, we constructed a physical map by screening and sequencing Korean Native Pig (KNP) BAC clones on the syntenic region between 65 Mb and 85 Mb of human chromosome 1. To fill the gaps in KNP BAC contig map, we selected several CHORI-242 BAC clones from FPC map. A total of 18 Mb genomic sequence was obtained. A total of 114 genes including non-coding RNA genes were found on that genomic region. On that region, 3,556 SNPs were found and 199 SNPs with 100Kb intervals were tested for polymorphism. Approximately 70% of SNPs revealed polymorphic. In this study, 3 SNPs for BFT and 21 SNPs for IMF were shown highly significant. In addition, six individuals of Korean Native pig were sequenced at on round respectively with next generation sequencing method. A total of 8 Gb genomic sequences were obtained and covered 86% on the reference genome sequence of Build9. These data will be used for development of selective molecular markers for breeding.
To investigate the genetic characteristics of Korean and foreign dogs,the superior genetic studies were drafted for genetic comparisons. We constructed a full-length cDNA library of Korean Jindo dog for liver and brain tissues. A total of 20,000 EST sequences from liver and brain of Jindo dogs were determined based on the bioinformatic analysis from 141 significant gene expression inJindo dogs. 25 of novel specific gene were founded. In addition, the 42k canine gene expression chip showed the differential expression in the muscles of Jindo and shepherd dog, which includes a total of 71genes. To develop a genetic marker included SNP and MS, Korean native dog and foreign dog was used to validation with 973 of cSNP mining. Using the program ' R' , canine 22k SNP chip was surveyed to develop a genetic marker for canine hip dysplasia. And, the development of multiplex PCR using 27 microsatellite makers has a convenient PCR condition at the same time reducing the time and expense. Canine identification markers will be used as a complete patent application for the prevention of stray dogs.
The number of abandoned dogs is increasing with the worsening of the economy and the rising of feed value. It was becoming a serious social problem because of the disease transmission and destruction of natural ecosystems by abandoned dogs been wild animal. In order to solve these problems, companion dogs necessary to secure its own genetic information and to establish the systematic tracking system. Using multiplex-PCR method with 27 microsatellite marker (MS marker) divided 3 set, various alleles occurring to 6 dog breed (Labrador Retriever, German Shepherd, English Springer Spaniel, Belgian Malinois, Jindo Dog, PoongSan Dog) make use of markers to determine allele frequency and heterozygosity. MS marker FH2834 and FH2790 have only two allele and most were found in 13 alleles at FH3381 and FH3399. Average heterozygosity of MS marker is 0.534 and especially, heterozygosity represented the highest value of 0.765 at FH3381. So, it was recognized appropriate allele frequency for individual identification and paternity diagnosis in companion dogs. Using multiplex-PCR method with MS marker, various alleles occurring to dog breed make use of markers to determine individual identification and paternity diagnosis, traits associated biomarkers and breed-specific marker for faster, more accurate and ways to reduce the analysis cost. Based on this result, a scientific basis was established to the existing pedigree data by applying genetics additionally. Animal registration system is expected to be conducted nationwide in future. The method expects to very useful this system.
There have been studies about dog’s personality and behavior, which is helpful to breed dogs as guide or companion. In this study, a questionnaire was developed using 54 Jindo dogs, which considered ten items about aggressiveness and sociability. The scores were analyzed by principle component analysis (PCA), after accounting for four variables: age, gender, growing place, and coat-colors. Our results from the PCA indicated three principle components, which classified ‘aggressiveness’, ‘sociability’ and unknown factor. The four variables did not significantly affect aggressiveness (P> 0.05). However, there was a relationship between coat-color and sociability, i.e., the Jindo dogs with fawn color were more sociable than the white ones (P< 0.1).
The Jindo dog is a Korean natural monument and is recognized by the Fédération Cynologique Internationale. A prominent feature is the diverse coat color within the breed. To analyze the genetic basis of variation in the Jindo coat color, we sequenced the protein-coding regions of the melanocortin 1 receptor gene (MC1R). The MC1R coding sequence was determined from 154 dogs in five breeds (Jindo, Labrador Retriever, English Springer Spaniel, Belgian Malinois, and German Shepherd). To confirm the genetic structure of sampled populations, we tested for Hardy-Weinberg equilibrium (HWE) and computed Fst. The sample populations did not significantly deviate from HWE. Fst was 0.02 between white and fawn Jindo dogs; this was lower than Fst between breeds. Six single nucleotide polymorphisms (SNPs) were detected in the MC1R coding region. Among the six SNPs, five were non-synonymous (S90G, T105A, Q159P, M264V, and R306ter) and one was synonymous SNP (Y298Y). From the SNPs, we predicted four haplotypes (H1, H2, H3, and H4) for Jindo MC1R. Jindo dogs had different haplotypes corresponding to different coat colors. H1 was frequently observed in white Jindo dogs with an odds ratio of 5.03 (95% CI: 2.27-11.18, p< 0.0001), whereas H2 and H4 were observed only in fawn Jindo dogs. Our findings indicate that SNP haplotype can influence coat color. Knowledge of MC1R haplotypes can help discriminate white and fawn coats in Jindo dogs. We hope this report will trigger more research into the genetics of this traditional Korean dog and will be a reference for dogs of Asian origin. Also, our results will provide a useful genetic marker for Jindo dog breeders who have selected for specific colors.
We generated 16,993 expressed sequence tags (ESTs) from two libraries containing full-length cDNAs from the brain and liver of the Korean Jindo dog. An additional 365,909 ESTs from other dog breeds were identified from the NCBI dbEST database, and all ESTs were clustered into 28,514 consensus sequences using StackPack. We selected the 7,305 consensus sequences that could be assembled from at least five ESTs and estimated that 12,533 high-quality single nucleotide polymorphisms (SNPs) were present in 97,835 putative SNPs from the 7,305 consensus sequences. We identified 58 Jindo dog-specific SNPs in comparison to other breeds and predicted seven synonymous SNPs and ten non-synonymous SNPs. Using PolyPhen, a program that predicts changes in protein structure and potential effects on protein function caused by amino acid substitutions, three of the non-synonymous SNPs were predicted to result in changes in protein function for proteins expressed by three different genes (TUSC3, ITIH2, and NAT2).

목차 Contents

• 표 지 ... 1
• 제출문 ... 2
• 요 약 문 ... 3
• SUMMARY ... 4
• 목 차 ... 7
• 제 1 장 서 론 ... 8
• 제 2 장 국내외 기술개발 현황 ... 10
• 제 1 절 : 세계적 수준 ... 10
• 제 2 절 : 국내수준 ... 11
• 제 3 절 : 국내외의 연구현황 ... 11
• 제 3 장 연구개발수행 내용 및 결과 ... 13
• 제1절 한국재래돼지 표준 유전체 지도 작성 ... 13
• 제2절 돼지 6번 염색체 q32-34 영역의 기준 유전체 서열 작성 ... 13
• 제3절 지방형질 QTL 영역 유전자 지도 작성 ... 14
• 제4절 돼지 유전체 해독 국제 컨소시엄(SGSC) 활동 ... 16
• 제5절 개의 초위성체마커에의한 유전적 근연관계분석 ... 16
• 제6절 개 경제형질관련 SNP 탐색 및 유전자마커 개발 ... 19
• 제 7절 진도개 간 및 뇌의 dbEST 구축 ... 23
• 제 8절. 서열데이터 수집 및 재가공 ... 24
• 제 4 장 연구개발 목표 달성도 및 대외 기여도 ... 32
• 제1절 목표대비 대외달성도 ... 32
• 제2절 정량적 성과 ... 33
• 제 5 장 연구개발결과의 활용계획 ... 41
• 제 1절 돼지유전체의 추가연구 필요성 ... 41
• 제 2절 개 유전체의 추가연구 필요성 ... 41
• 제 3절 가축생물정보의 추가연구의 필요성 ... 41
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 42
• 제 7 장 기타 중요 변동사항 ... 43
• 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 44
• 제 9 장 참고문헌 ... 45
• 끝페이지 ... 51