MOLECULAR ANALYSIS OF ORIGIN OF JAPANESE DOMESTIC DOGS BY ANCIENT DNA

从绳纹、弥生、古fun、鄂霍次克和镰仓时期的145个古代犬类样本（主要是骨头）中确定了日本犬类考古遗骸的古代线粒体DNA （mtDNA）序列。从独立制备的DNA提取物中扩增出线粒体控制区（198bp）的高度可变序列至少两次，并将来自古代样本的序列与来自178只现代狗的*单倍型进行比较。从74份古犬样本中扩增出198bp的古代mtDNA，并将序列划分为19个单倍型，其中包括现代犬中发现的5个现代单倍型（M1、M2、M5、M10和M11）。通过系统发育分析，在现代犬对照区划分出3个主要聚类（CL1 ~ CL3）；所有的古代狗都属于CL1集群。北海道犬的起源被认为是来自南方犬群，而不是来自北方犬群。冲绳古代犬系通过贸易活动从欧亚大陆演化而来，古苏时期犬系与现代琉球犬系具有相同的单倍型。测定了日本狼的mtDNA，并对日本狼、犬和狼的遗传关系进行了研究。日本狼与犬、狼的mtDNA系统发育分析表明，日本狼与来自欧亚大陆的犬、狼有亲缘关系。为了加强犬类mtDNA数据库，从365只犬中扩增出600bp的mtDNA d -环区，并测定mtDNA序列。将600bp序列的多样性划分为64个单倍型，其中新发现的46个单倍型，并将其分为I ~ IV 4个聚类。

Sequences of ancient mitochondrial DNA (mtDNA) from archaeological remains of Japanese dogs were determined from 145 ancient dog samples (mainly bones) from the Jomon, Yayoi, Kofun, Okhotsk and the kamakura Periods. Highly variable sequences of the mitochondrial control region (198bp) were amplified at least twice from independently prepared DNA extracts, and those from ancient samples were compared with * haplotypes from 178 modern dogs. The 198bp ancient mtDNA was amplified from 74 ancient dog samples, and the sequences were classified into 19 haplotypes comprising five modern haplotypes (M1, M2, M5, M10 and M11) identified in modern dogs. Three major clusters (CL1 to CL3) were distinguished within the modern dog control region using phylogenetic analysis; all ancient dogs belonged to the CL1 cluster. Origin of Hokkaido dogs was considered to be from southern dog population, but not from northern dog population. Ancient dogs in Okinawa Islands was derived from Eurasian continent through trade activity, and dogs in Gusuku Period had the same haplotypes to modern Ryukyu dog populations. The mtDNA from Japanese wolf was also determined, the genetical relationship among Japanese wolf, dogs and wolves were investigated. The phylogenetic analysis of mtDNA from Japanese wolf with those from dogs and wolves indicated that Japanese wolf genetically related to dogs and wolves from Eurasian continent.To reinforce the canine mtDNA database, 600bp of mtDNA D-loop region was amplified from 365 dogs and determined for mtDNA sequence. The diversity of the 600bp sequences was classified into 64 haplotypes, including 46 newly discovered haplotypes, and the haplotypes were grouped into four clusters I to IV.