報告書番号

U00016

タイトル（和文）

DNA多型を利用したノウサギの個体識別

タイトル（英文）

Individual Identification of Japanese Hare Using DNA Polymorphism

概要 （図表や脚注は「報告書全文」に掲載しております）

ノウサギについて、アナウサギで同定されているSTR（Short Tandem Repeat）多型検出用プライマーを用いてSTRが検出できるか検討を行った。PCR反応条件を検討した結果、10種類のSTRのうち7種類でノウサギでもPCRの増幅が可能であった。増幅したPCR産物について塩基配列を決定した結果、いずれもアナウサギの各STRの塩基配列と高い相同性を示し、反復配列を有することが確かめられた。15頭のノウサギについて7種類のSTRを用いて多型解析を行ったところ、どの個体同士でも同一の多型パターンを示す組み合わせが無く、全個体を識別することが可能であった。解析した15頭の中の血縁関係にも遺伝的に矛盾はなく、親子や兄弟間の識別も可能であったことから、使用した7種類のSTRの組み合わせによる多型解析は、ノウサギの個体識別に有用であることが明らかとなった。

概要 （英文）

Short tandem repeats (STR) are highly polymorphic and short nucleotide repeating units scattered randomly throughout the genome. They are therefore promising genetic markers which can use for the identification of individuals, for the determination of kinship structures and for the estimation of population size. In this study, we describe analysis of STR polymorphism in Japanese hare (Lepus brachyurus angustidens), which is important as the prey of raptorial birds. To examine whether STR loci isolated from European wild rabbit (Oryctolagus cuniculus) are suitable for the identification of individuals in Japanese hare, ten rabbit STRs were amplified by polymerase chain reaction (PCR). After optimization of various PCR parameters such as annealing temperature and magnesium chloride concentration, seven of the ten rabbit STRs were amplified successfully in Japanese hare. Sequence analysis confirmed that the seven STR loci amplified in Japanese hare contained tandemly repeated sequences and showed significant homology to the sequences of European wild rabbit STRs. A total of fifteen DNA samples, which extracted from blood, muscular tissues and hair, were genotyped with the seven STR loci. As there was no combination that shows the equal pattern of the STR genotypes in any of samples, these STR typing enabled all fifteen Japanese hares to be distinguished from each other. Mendelian inheritance of the observed polymorphism was confirmed by family studies. These results suggested that the seven STRs are sufficiently polymorphic to be useful as markers in genetic studies in hares.

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