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Spermophilus dauricus, a member of the subfamily Sciurinae and family Scuidae, is an endemic rodent of North China.Spermophilus is one of the most diverse genera including 6 species of squirrels, occurring from tropical to high-latitude regions in China.Extensive researches of these species have been conducted on their behavior, ecology, and reproductive biology.Yet consensus of taxonomic status of this group has not been reached.In this study, S.dauricus tissue samples were collected from Tianzhu, Gansu Province for extracting DNA.We sequenced 1,140 bp of Cyt b gene from PCR products.Clustal X1.84, MEGA 6.0, DnaSP 5.0 and other bioinformatics softwares were used for sequence analysis.The results showed that Cyt b gene have over 116 polymorphic information sites.Population polymorphisms are result from transition and transversion.T, C, A, G nucleotide contents of Cyt b gene were 33.7%, 25.9%, 27.6% and 12.9%, respectively.Based on genetic distance of Kimura two-parameter, intraspecific genetic distance between large ground squirrel (S.major) and Xinjiang ground squirrel (S.fulvus) is the smallest (0.034), and that between Caucasian dauricus (S.musicus) and ground squirrels (S.dauricus) is the largest (0.172).The genetic distance between populations of S.dauricus is 0.140, suggesting large population differentiation, N J-based phylogenetic tree confirms the traditional taxonomy.Our findings provide baseline information for further studies in phylogenetics and population genetics of Spermophilus.