미토콘드리아 DNA CYTB 유전자 서열에 대한 분자 계통과 PCR-RFLP 반수체형에 근거한 제주재래돼지의 모계 기원 Maternal Origins of the Jeju Native Pig Inferred from PCR-RFLP Haplotypes and Molecular Phylogeny for Mitochondrial DNA CYTB Gene Sequences원문보기
제주재래돼지의 모계 혈통에 대한 보다 명확한 이해를 얻기 위해, 본 연구에서는 제주재래돼지의 미토콘드리아 DNA (mtDNA) CYTB 유전자를 분석하고 이를 타 품종들에서 얻은 결과들과 비교하였다. 제주재래돼지를 포함한 돼지 6 품종에서 PCR-RFLP 분석을 수행하였고, RFLP 양상은 돼지 품종들을 뚜렷하게 구분되는 두 가지 반수체형(mtCYTB1 and mtCYTB2)으로 분리시켰다. 제주재래돼지 CYTB 서열들은 계통수 상에서 유럽과 아시아품종 cluster에서 모두 발견되었다. 제주재래돼지 CYTB들 중에서 J2 group은 중국재래돼지품종들과 근연이면서 아시아 고유 돼지 계통들과 함께 출현하였으며, 다른 한 group인 J1에 해당하는 서열들은 유럽돼지 계통들과 함께 위치하였고, 아시아 품종들보다는 스페인의 Iberian 재래돼지들과 근연인 것으로 확인되었다. 이 결과들은 현재 제주도에서 사육되고 있는 제주재래돼지 품종의 모계 기원은 크게 아시아계 돼지와 유럽계 돼지인 것으로 추정됨을 보여준다. 따라서 본 연구결과들은 제주재래돼지 집단은 과거에 가축화된 아시아 고유 돼지품종들과 공통 선조를 공유하고, 또한 20세기에 유입된 유럽계 돼지 품종들도 현재의 집단 형성에 기여한 것임을 시사하고 있다.
제주재래돼지의 모계 혈통에 대한 보다 명확한 이해를 얻기 위해, 본 연구에서는 제주재래돼지의 미토콘드리아 DNA (mtDNA) CYTB 유전자를 분석하고 이를 타 품종들에서 얻은 결과들과 비교하였다. 제주재래돼지를 포함한 돼지 6 품종에서 PCR-RFLP 분석을 수행하였고, RFLP 양상은 돼지 품종들을 뚜렷하게 구분되는 두 가지 반수체형(mtCYTB1 and mtCYTB2)으로 분리시켰다. 제주재래돼지 CYTB 서열들은 계통수 상에서 유럽과 아시아품종 cluster에서 모두 발견되었다. 제주재래돼지 CYTB들 중에서 J2 group은 중국재래돼지품종들과 근연이면서 아시아 고유 돼지 계통들과 함께 출현하였으며, 다른 한 group인 J1에 해당하는 서열들은 유럽돼지 계통들과 함께 위치하였고, 아시아 품종들보다는 스페인의 Iberian 재래돼지들과 근연인 것으로 확인되었다. 이 결과들은 현재 제주도에서 사육되고 있는 제주재래돼지 품종의 모계 기원은 크게 아시아계 돼지와 유럽계 돼지인 것으로 추정됨을 보여준다. 따라서 본 연구결과들은 제주재래돼지 집단은 과거에 가축화된 아시아 고유 돼지품종들과 공통 선조를 공유하고, 또한 20세기에 유입된 유럽계 돼지 품종들도 현재의 집단 형성에 기여한 것임을 시사하고 있다.
In an effort to gain greater understanding of the maternal lineages of the Jeju native pig (JNP), we analyzed the mitochondrial DNA (mtDNA) CYTB gene and compared it with those of other pig breeds. PCR-RFLP analysis was conducted with six pig breeds including JNP, and then the RFLP patterns allowed ...
In an effort to gain greater understanding of the maternal lineages of the Jeju native pig (JNP), we analyzed the mitochondrial DNA (mtDNA) CYTB gene and compared it with those of other pig breeds. PCR-RFLP analysis was conducted with six pig breeds including JNP, and then the RFLP patterns allowed for the separation of the pig breeds into two distinct haplotypes (mtCYTB1 and mtCYTB2). The JNP CYTB sequences were detected in both the European and Asian breed clusters on the phylogenetic tree. The J2 group was sorted with the indigenous cluster of Asian pig lineages and was related closely to Chinese native pig breeds, but a second group, J1, was sorted with the European pig lineages and appeared to be related to Spanish Iberian native pigs, rather than to Asian breeds. These results indicate that the JNP currently raised on Jeju Island have two major maternal origins estimated in Asian and European pigs. We concluded that the JNP that share a common lineage with indigenous Asian pigs were domesticated in the distant past, originating from pigs that were already being raised elsewhere at that time, and that the European pig breeds introduced in the twentieth century have also contributed to the formation of this pig population.
In an effort to gain greater understanding of the maternal lineages of the Jeju native pig (JNP), we analyzed the mitochondrial DNA (mtDNA) CYTB gene and compared it with those of other pig breeds. PCR-RFLP analysis was conducted with six pig breeds including JNP, and then the RFLP patterns allowed for the separation of the pig breeds into two distinct haplotypes (mtCYTB1 and mtCYTB2). The JNP CYTB sequences were detected in both the European and Asian breed clusters on the phylogenetic tree. The J2 group was sorted with the indigenous cluster of Asian pig lineages and was related closely to Chinese native pig breeds, but a second group, J1, was sorted with the European pig lineages and appeared to be related to Spanish Iberian native pigs, rather than to Asian breeds. These results indicate that the JNP currently raised on Jeju Island have two major maternal origins estimated in Asian and European pigs. We concluded that the JNP that share a common lineage with indigenous Asian pigs were domesticated in the distant past, originating from pigs that were already being raised elsewhere at that time, and that the European pig breeds introduced in the twentieth century have also contributed to the formation of this pig population.
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문제 정의
However, the domestication process and the origin of present JNP remain to be clearly elucidated, and the phylogenetic relationship of JNP with other pig populations has yet to be definitively established. In an effort to shed light on these remaining questions, in this study we have assessed the sequences and haplotypes of the mtDNA CYTB gene.
제안 방법
In order to extrapolate the phylogenetic relationship of JNPs from the CYTB gene sequences, we obtained 88 complete CYTB sequences from the GenBank database using a BLAST search of our sequences. However, we did not apply the sequences from the wild boar for the phylogenetic analysis, in order to prevent misunderstandings regarding the genetic source of JNP.
In order to investigate the maternal origin of JNPs, complete sequences of mtDNA CYTB gene from JNPs and the other five pig breeds, Berkshire, Duroc, Hampshire, Landrace and Large White, were determined. Additionally, to scan the genetic diversity of each pig population, PCR-RFLP analysis was also conducted.
TA Cloning Kit (Invitrogen, Carlsbad, CA, USA). Nucleotide sequencing was conducted using a DYEnamic ET-Dye Terminator Kit (GE Healthcare) and separated on a MegaBace1000 automated DNA sequencer (GE Healthcare). The nucleotide sequences acquired in this study have been submitted to the GenBank database (GenBank accession nos.
대상 데이터
Blood samples were obtained from 65 JNP from the Subtropical Animal Experiment Station (SAES), National Institute of Animal Science, Rural Development Administration and Institute for Livestock Promotion (ILP), Jeju-do in South Korea. 62 Berkshire and 10 Hampshire pigs were generously provided from pig farms in Jeju-do and Gyeongsangnam-do.
Nucleotide sequencing was conducted using a DYEnamic ET-Dye Terminator Kit (GE Healthcare) and separated on a MegaBace1000 automated DNA sequencer (GE Healthcare). The nucleotide sequences acquired in this study have been submitted to the GenBank database (GenBank accession nos. AY830173- AY830188).
이론/모형
AJ314554) for out-group rooting. Phylogenetic analysis was conducted using NEIGHBOR in the PHYLIP program package [5]. The neighbor-joining (NJ) tree was constructed on the basis of pairwise genetic distances calculated via the two-parameter method [16].
Phylogenetic analysis was conducted using NEIGHBOR in the PHYLIP program package [5]. The neighbor-joining (NJ) tree was constructed on the basis of pairwise genetic distances calculated via the two-parameter method [16].
성능/효과
In other pig breeds, two restriction patterns were also detected, which with the exception of Landrace and Large White pigs, were similar to those from JNP (Table 2). Combining the restriction patterns, two kinds of CYTB haplotypes, mtCYTB1 and mtCYTB2, were detected (Table 3). Both haplotypes were found in the four pig breeds-Berkshire, Hampshire, Duroc, and JNP-however, the Large White pig ha the mtCYTB2 haplotype, and the Landrace pigs harbor the mtCYTB1 haplotype.
후속연구
However, there is still not sufficient data to adequately evaluate the relationship and evolutionary origins of Korean native pigs, including JNPs. In order to establish clearly the relationship between the domestication histories of native pigs and wild boars in Korea, and to clearly characterize the progenitor population and more accurately elucidate the phylogenetic relationships of the extant wild boars, detailed future studies are recommended, with an expanded sample population of present and ancient pigs from Korea and East Asia.
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