학술논문
염소의 DNA 개체식별을 위한 초위성체 마커 개발
Development of Microsatellite Markers for DNA Individual Identification of goats
Development of Microsatellite Markers for DNA Individual Identification of goats
Document Type
Article
Source
한국산학기술학회논문지, 24(11), pp.409-417 Nov, 2023
Subject
Language
한국어
ISSN
2288-4688
1975-4701
1975-4701
Abstract
본 연구의 목적은 염소 및 염소고기의 부정 유통을 근절하고 과학적인 혈통관리를 통한 염소 개량 효과의 극대화를 위하여 염소 개체식별 및 친자감정 DNA 감식법 개발하는 것이다. 염소의 대립유전자 판별 오류를 줄이고 분석 결과의 신뢰도와 정확도를 높이기 위해 3반복과 4반복 microsatellite 좌위들로 이루어진 새로운 염소 개체식별 유전자 마커 15종(KCH004, KCH006, KCH015 KCH027, KCH047, KCH067, KCH086, KCH114, KCH117, KCH121, KCH122, KCH134, KCH153, KCH165, KCH212)을 개발하였다. 선발된 15개의 좌위를 이용하여 염소 96두에 대하여 다중 PCR(multiplex PCR) 초위성체 유전자형 결정을 실시한 결과, 총 54개에 대립유전자가 발견되었으며 좌위별로평균 3.60개의 대립유전자를 가지는 것으로 확인되었다. 마커의 다형성과 정보력의 척도인 PIC(Polymorphism Information Contents) 값은 0.090(KCH153)~0.545(KCH121)로 나타났으며 KCH006, KCH015, KCH114, KCH121 좌위들은 PIC 0.35 이상이었고 전체 15종 마커의 평균PIC 값은 0.28을 가지는 것으로 확인되어 개체식별마커로서 다형성이 있음이 검증되었다. 개발한 마커를 활용하여 재래흑염소 (당진, 장수, 통영 계통)와 외국 5품종(알파인, 보어, 누비안, 자아넨, 토겐부르크)의 유전적 특성을 분석하였으며, 총 6품종 8계통 집단과 각각의 유전자 마커의이형접합율(Heterozygosity)을 측정하였다. 기대 이형접합율은 0.167(누비안)~0.467(보어) 범위 내로, 재래흑염소는0.393(당진), 0.415(통영), 0.437(장수)의 값을 가지는 것으로 나타났다. 재래흑염소(당진, 장수, 통영)와 외국 5품종들간의 유연관계 분석은 특정 대립유전자 빈도를 근거로 한 유전적 거리의 추정으로 이루어졌다. 재래흑염소와 보어 집단간 유전적 거리가 가장 가깝고 자아넨과 가장 먼 것으로 나타났다. 염소의 유전적 분화 양상을 확인함으로써 개발된15종의 마커가 개체식별, 친자감정의 유전자 감식뿐만 아니라 염소 유전적 다양성 및 유연관계 분석에 활용될 수 있을것으로 사료된다.
The purpose of this study was to develop individual goat identification and paternity DNA testing methods to eradicate the illegal distribution of goats and goat meat and maximize the effects of goat improvement through scientific pedigree management. To reduce goat allele discrimination errors and increase the reliability and accuracy of the analysis, 15 new goat individual identification genetic markers consisting of 3- and 4-repeat microsatellite loci (KCH004, KCH006, KCH015 KCH027, KCH047, KCH067, KCH086, KCH114), KCH117, KCH121, KCH122, KCH134, KCH153, KCH165, KCH212) were developed. As a result of performing muliplex polymerase chain reaction (PCR) microsatellite genotyping on 96 goats using the selected 15 loci, a total of 54 alleles were found, and it was confirmed that each locus had an average of 3.60 alleles. The polymorphism information content (PIC) value, which is a measure of polymorphism and information power of markers, ranged from 0.090 (KCH153) to 0.545 (KCH121), and loci KCH006, KCH015, KCH114, and KCH121 had a PIC of 0.35 or higher. The average PIC value of all 15 markers was confirmed to be 0.28, and it was verified that there was a polymorphism as an individual identification marker. Using the developed markers, the genetic characteristics of native black goats (Dangjin, Jangsu, and Tongyeong lines) and five foreign breeds (Alpine, Boer, Nubian, Saanen, and Toggenburg) were analyzed. The heterozygosity of the genetic markers was measured. The expected heterozygous ratio was found to be within the range of 0.167 (Nubian) to 0.467 (Boer), and the native black goat had values of 0.393 (Dangjin), 0.415 (Tongyeong), and 0.437 (Jangsu). The analysis of the affinity relationship between native black goats (Dangjin, Jangsu, and Tongyeong) and the five foreign breeds was performed by estimating the genetic distance based on specific allele frequencies. It was found that the genetic distance between the native black goat and the Boer population was the closest and it was farthest from Saanen. It is believed that the 15 markers developed by confirming the genetic differentiation patterns of goats can be used not only for individual identification and paternity genetic identification but also for goat genetic diversity and related relationship analysis.
The purpose of this study was to develop individual goat identification and paternity DNA testing methods to eradicate the illegal distribution of goats and goat meat and maximize the effects of goat improvement through scientific pedigree management. To reduce goat allele discrimination errors and increase the reliability and accuracy of the analysis, 15 new goat individual identification genetic markers consisting of 3- and 4-repeat microsatellite loci (KCH004, KCH006, KCH015 KCH027, KCH047, KCH067, KCH086, KCH114), KCH117, KCH121, KCH122, KCH134, KCH153, KCH165, KCH212) were developed. As a result of performing muliplex polymerase chain reaction (PCR) microsatellite genotyping on 96 goats using the selected 15 loci, a total of 54 alleles were found, and it was confirmed that each locus had an average of 3.60 alleles. The polymorphism information content (PIC) value, which is a measure of polymorphism and information power of markers, ranged from 0.090 (KCH153) to 0.545 (KCH121), and loci KCH006, KCH015, KCH114, and KCH121 had a PIC of 0.35 or higher. The average PIC value of all 15 markers was confirmed to be 0.28, and it was verified that there was a polymorphism as an individual identification marker. Using the developed markers, the genetic characteristics of native black goats (Dangjin, Jangsu, and Tongyeong lines) and five foreign breeds (Alpine, Boer, Nubian, Saanen, and Toggenburg) were analyzed. The heterozygosity of the genetic markers was measured. The expected heterozygous ratio was found to be within the range of 0.167 (Nubian) to 0.467 (Boer), and the native black goat had values of 0.393 (Dangjin), 0.415 (Tongyeong), and 0.437 (Jangsu). The analysis of the affinity relationship between native black goats (Dangjin, Jangsu, and Tongyeong) and the five foreign breeds was performed by estimating the genetic distance based on specific allele frequencies. It was found that the genetic distance between the native black goat and the Boer population was the closest and it was farthest from Saanen. It is believed that the 15 markers developed by confirming the genetic differentiation patterns of goats can be used not only for individual identification and paternity genetic identification but also for goat genetic diversity and related relationship analysis.