Phylogeny of extinct wolves based on nucleotide saguence

基于核苷酸序列的已灭绝狼的系统发育

• 批准号: 05640787
• 负责人: TAKENAKA Osamu
• 金额: $ 1.34万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05640787/
• 关键词: Jamainu; Japanese wolf; Ezo-wolf; Multivariate Analysis; DNA; Molecular Phylogeny; PCR; Mitochondria

The present study aimed morphological examination of skeletal and skin specimens of extinct Japanese wolves and wild dogs which have been kept in various institutions. We attempted to determine thenucleotide sequence of part of mitrochonrial DNA using those pelage samples.We could get morphological data and a piece of skin samples from the specimens kept at Leiden museum in the Netherlands. We could also prepare the skin samples from, Faculty of Agriculture of Hokkaido University, National Science Museum, Tokyo. Neary 40 morphological data and 9 skin samples were collected.Multivariate analyzes of the measurement revealed wolves in Asian continent are close to Ezo-wolves in Hokkaido, next is domestic dogs and these species are rather distant relationships with the Japanese wolves.Polymerase chain reaction using DNA from skin samples as template and primer set for cytochrome b gene of mitochondrial DNA could amplify DNA fragment from two specimens out of 5 examined. We could not amplify any DNA for other 3 samples. Some pale brownish substance (s) may inhibit DNA polymerase. Computer analyzes of the sequences revealed that one is cytochrome b gene of unkown eukaryote and the other is cyto-chrome b of human. We will continue to purify the DNA samples before PCR by various methods.

本研究的目的是形态学检查已灭绝的日本狼和野狗的骨骼和皮肤标本保存在不同的机构。我们试图利用这些样本确定部分线粒体DNA的核苷酸序列。我们可以从荷兰莱顿博物馆保存的标本中获得形态学数据和一块皮肤样本。我们也可以从北海道大学农学院，国立科学博物馆，东京准备皮肤样本。收集了近40份形态学数据和9份皮肤样本。多变量分析表明，亚洲大陆的狼与北海道的野狼关系密切，其次是家犬，这些物种与日本狼的关系较远。用皮肤样本DNA作为模板和线粒体DNA细胞色素b基因引物，聚合酶链反应可扩增5个样本中的2个样本的DNA片段。其他3个样本我们无法扩增DNA。一些淡棕色物质可能抑制DNA聚合酶。经计算机分析，这两个基因序列一个是未知真核生物的细胞色素b基因，另一个是人类细胞色素b基因。我们将继续用各种方法纯化PCR前的DNA样本。

项目成果

H.Kakoi: "Divergence between the Anoas of Sulawesi and the Asiatic Water buffaloes,infevred from their complete amino and Seguences of hemoglobin B chains." Z.zeol.syst.Evolut.-forsch. 32. 1-10 (1994)

H.Kakoi：“苏拉威西阿努阿牛和亚洲水牛之间的差异，从它们的完整氨基酸和血红蛋白 B 链序列推断出来。”

O.Takenaka: "Molecular evolution of the genus Macaca (in Japanese) ." Iden. 48. 30-34 (1994)

O.Takenaka：“猕猴属的分子进化（日语）。”

H.Suzuki: "Phylogenetic relationships among Hnmo sapiens and related species Dased on restriction site variations in rDNA spacers." Biochem,Genet.32. 257-269 (1994)

H.Suzuki：“基于 rDNA 间隔区限制性位点变异，智人和相关物种之间的系统发育关系。”

竹中修: "分子進化からみたマカカ属の種分化" 遺伝. 48. 30-34 (1994)

Osamu Takenaka：“从分子进化角度观察猕猴属的物种形成”，《遗传学》48. 30-34 (1994)。

T.Tanaka: "Estimation of phylogenetic relationships among Japanese brown frogs from Mitochondrial oxto chrome b gene" Zool.Sui.11. 753-757 (1994)

T.Tanaka：“从线粒体牛铬b基因估计日本褐蛙的系统发育关系”Zool.Sui.11。