16S rRNA Gene Sequence Analysis of Snow Leopard, Gray Wolf, Horse and Bactrian Camel in Mongolia

Munkhtuul Tsogtgerel, Munkhtogtokh Baljijjnyam, Nansalmaa Suren, Lkhagvasuren Sodnom
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Abstract

In this study, mitochondrial 16S rRNA sequences of snow leopard, gray wolf, domestic horse and Bactrian camel inhabited or domesticated in Mongolian territory were obtained by using polymerase chain reaction (PCR) based on universal primers for 16S rRNA (F-5'-AACGAGCCTGGTGATA-3' and R-5'-CTCCGGTCTGAACTCAGATCACGTA-3’). The 16S rRNA sequence was 1,048 bp to 1,086 bp in length, and each sequence was compared to other related species (Felidae, Camelidae, Equidae and Canidae) by using NCBI Basic Local Alignment Search Tool (BLAST). Results showed that sequences were highly similar to sequences in GenBank database (93%-99%). Then phylogenetic analysis was performed based on about 1,100 bp sequence of 16S rRNA for Panthera uncia, Canis lupus, Equus caballus, Camelus bactrianus and other related species. The result revealed that P. uncia and P. leo were sister species, C. bactrianus and C. ferus were more closely related species, and wolf and dog were the almost similar species. This finding could be important for designing species specific primers for PCR based analysis of animal species identification and forensic veterinary medicine.
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蒙古雪豹、灰狼、马和双峰驼16S rRNA基因序列分析
本研究采用基于16S rRNA通用引物(F-5′-AACGAGCCTGGTGATA-3′和R-5′-CTCCGGTCTGAACTCAGATCACGTA-3′)的聚合酶链式反应(PCR)方法,获得蒙古境内居住或驯化的雪豹、灰狼、家马和双峰驼的线粒体16S rRNA序列。16S rRNA序列长度为1048 ~ 1086 bp,利用NCBI Basic Local Alignment Search Tool (BLAST)将每个序列与其他近缘种(Felidae、Camelidae、Equidae和Canidae)进行比对。结果表明,该序列与GenBank数据库中的序列高度相似(93% ~ 99%)。然后,基于近缘种Panthera uncia、Canis lupus、Equus caballus、Camelus bactrianus等近缘种16S rRNA约1100 bp序列进行系统发育分析。结果表明,uncia与leo为姊妹种,C. bactrianus与C. ferus亲缘关系较近,狼与狗的亲缘关系较近。这一发现对设计物种特异性引物用于动物物种鉴定和法医兽医学分析具有重要意义。
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