长爪沙鼠Tim-1基因部分序列克隆及表达分析

Molecular cloning of partial sequence and expression analysis of Tim-1 gene in Meriones unguiculatus

根据美国国立生物技术信息中心(NCBI)中基因库(GenBank)里查询到已登录的小鼠、大鼠的Tim-1mRNA序列,通过同源比较,设计引物,首次对长爪沙鼠Tim-1基因部分编码序列进行分子克隆.经过PCR扩增,获得长爪沙鼠Tim-1基因243bp部分编码序列,经测序后登录GenBank(JN628997),发现该序列与小鼠、大鼠Tim-1相应部分有80%以上的相似性.利用所克隆的序列设计引物,建立荧光定量PCR方法检测长爪沙鼠Tim-1基因在不同组织中表达情况.结果显示,长爪沙鼠Tim-1基因在不同组织中表达差异性较大,其中在肾脏组织中表达水平高于其他组织,在肌肉、心脏、小肠、脾脏、肝脏、肺组织中表达均较低.

The present study was using the accessed sequences of Mus musculus,Rattus norvegicus Tim-1 mRNA that checked from GenBank of NCBI,through multiple alignment,homologized primers were designed based on them.This was the first time to clone Tim-1 gene from Meriones unguiculatus.By PCR amplification,243 bp（GenBank Accession： JN628997） partial coding sequence of M.unguiculatus Tim-1 gene were obtained.Sequence alignment revealed that the similarity of 243 bp segmnent in M.unguiculatus Tim-1 gene was over 80% compared with corresponding sequences in Mus musculus and Rattus norvegicus.The Tim-1 expression levels in multiple tissues were determined by real time quantitative PCR.The results showed that the levels of Tim-1 gene expression in the kidney is constantly higher than that in other tissues.Relatively lower abundance of Tim-1 transcripts can be detected in the muscle,heart,small intestine,spleen,liver,and the lung.