RAPD기법을 이용한 축우의 유전적 다형성과 유사도 분석

Analysis of Genetic Polymorphisms and Similarity Using Random Amplified Polymorphic DNAs in Cattle

본 연구는 축우 품종의 유전적 다형성을 분석하여 분자유전수준에서 축우의 품종간 판별과 유전적 유사성을 구명하고자 Holstein종과 한우, Charolais종 및 한우 교잡종의 혈액에서 DNA를 추출하여, PCR(polymerase chain reaction) 기법에 의한 RAPD(random amplified polymorphic DNAs)분석을 실시하고, 소 품종간의 유사도 지수를 추정한 후 UPGMA(unweighted pair-group method using average)방법으로 유사도를 추정하였으며 그 결과를 요약하면 다음과 같다. 1. 축우 품종들로부터 추출된 genomic DNA를 1.5% agarose gel에서 전기영동한 결과, 12.2kb 이상의 단일밴드로 나타났으며, genomic DNA의 순도는 DNA 흡광도 $A_{260}$과 $A_{280}$의 비율로 측정한 결과 그 비율은 1.75~2.10이었다. 2. RAPD기법에 의한 축우 품종의 유전적 다형성을 분석한 결과, 품종판별을 위한 가장 유용한 random primer는 5'-GAC CGC TTG T-3'인 것으로 판명되었다. 3. Primer 부착 온도에 따른 band 양상에 있어서 $33.0^{\circ}C$인 경우는 약 340bp에서 Holstein종과 Charolais종에서는 band가 출현되었으나, 한우와 한우 교잡종에서는 밴드가 나타나지 않았으며, $37.5^{\circ}C$의 경우에는 340bp에서 Holstein종과 한우에서는 밴드가 나타났으나, Charolais종과 한우 교잡종에서는 band가 출현되지 않아 이들 밴드의 유무가 소 품종판별을 위한 유용한 유전적 표지로서의 이용이 가능할 것으로 사료되었다. 4. 축우 품종의 유사도 지수에 있어서는 primer 부착온도를 $33.0^{\circ}C$와 $37.5^{\circ}C$을 종합하였을때 Holstein종과 한우간에는 0.666~0.777이었으며, Charolais종간에는 0.615~0.666이었고, 한우와 Charolais종간에는 0.400~0.461로 다소 낮은 수치를 보였으나, 한우와 한우 교잡종간에는 0.857~0.888로 대체로 높은 계수를 보였다. 5. 이상의 결과를 종합하여 보면 RAPD기법에 의하여 random primer 5'-GAC CGC TTG T-3'으로 축우 품종의 판별이 가능하며, UPGMA에 의한 축우 품종의 유사도는 Holstein종과 한우 및 Charolais종간 보다는 한우와 한우 교잡종 및 Charolais종간의 유전적 유사도가 높은 것으로 나타났다.

This study was carried out to investigate the difference and genetic similarity at the level of molecular genetics. Genomic DNA was extracted from blood of Holstein, Korean cattle, Charolais, and hybrid between Korean cattle and charolais and RAPD(random amplified polymorphic DNAs) was analyzed by PCR(polymerase chain reaction). After genetic similarity value from different breeds are analyzed, genetic similarity was estimated by UPGMA(unweighted pair-group method using average). The results obtained from this study can be summarized as follows: 1. When genomic DNA which was extracted from different breeds was subjected to electrophoresis on 1.5% agarose gel, bigger than 12.2kb was appeared. Ratio by absorbance of $A_{260}/A_{280}$ was 1.75~2.10, indicating that genomic DNA was quite pure for RAPD analysis. 2. Different band patterns by RAPD were appeared according to the breeds in cattle. The best primer used to distinguish Holstein from other breeds was 5'-GAC CGC TTG T-3'. 3. A 340bp fragment was amplified in $33.0^{\circ}C$ of annealing temperature for the Holstein and Charolais breeds, but any amplification was not occurred in this annealing temperature for Korean cattle and hybrid. In addition, a 340bp fragment was amplified in $37.5^{\circ}C$ of annealing temperature for the Holstein and Korean cattle, but any amplification was not occurred in this annealing temperature for Charolais and hybrid. For the reaction of PCR. $37.5^{\circ}C$ and $33.0^{\circ}C$ of annealing temperature was shown to be best for genetic marker identification from Holstein, Charolais, and hybrid between Korean cattle and Charolais. 4. When genetic similarity from different breeds are analyzed at the both temperature of $33.0^{\circ}C$ and $37.5^{\circ}C$, the genetic similarity value of Holstein and Korean cattle, Holstein and Charolais, Korean cattle and Charolais, and Korean cattle and hybrid were 0.666~0.777, 0.615~0.666, 0.400~0.461 and 0.857~0.888, respectively. 5. It could be concluded that different breeds are capable of distinguishing by RAPD used random primer 5'-GAC CGC TTG T-3', genetic similarity from different breeds was appeared the higher genetic similarity value of Korean cattle and Charolais than that of Holstein between Korean cattle and Charolais by UPGMA.