兔CCR10基因的克隆与序列分析

摘 要：【研究目的】克隆并分析兔CCR10基因；【方法】基于电子延伸序列，设计1对克隆引物，兔盲肠黏膜组织提取总RNA，进行RT-PCR，将PCR产物与pMD19-T载体连接后转化E.coli JM109感受态细胞、检测阳性克隆、测序并进行序列分析。【结果】克隆的兔CCR10 基因长为723bp，编码由241个氨基酸残基组成的CCR10前体蛋白，三级结构预测表明CCR10具有一个多克隆免疫球蛋白区（PIG-X）。克隆的兔CCR10基因与绵羊、人的同源性分别为89.6%、89.9%，推导的氨基酸序列与绵羊、人的同源性分别为94.2%、93.8%，结构特征与绵羊、人的相一致。【结论】克隆了兔CCR10基因并注册GenBank (Accession. EU348829)。

Molecular Cloning and Sequence Analysis of Oryctolagus cuniculus CCR10

Abstract 【Objective】To clone and analysis the Oryctolagus cuniculus CCR10. 【Method】A pair of cloning primers were designed based on the in silico sequence information, Total RNA was extracted from intestinum caecum mucosal tissue of Oryctolagus cuniculus and mRNA sequence was amplified by RT-PCR. The PCR products were ligated into the pMD19-T vector, and then transformed into competent cells of E.coli JM109. The positive clone was identified and the sequence was sequenced and analysis. 【Results】Oryctolagus cuniculus CCR10 consists of 723bp and encodes 241 amino acids . Tertiary structure predicts that CCR10 contains a PIG-X domain. Identity analysis showed that the CCR10 nucleotide sequence shared 89.6%、89.9% homology with that of Oryctolagus cuniculus、Ovis aries、Homo sapiens, the predicted peptide shared 94.2%、93.8% homology with that of Oryctolagus cuniculus、Ovis aries、Homo sapiens. revealing that the structural feature is consistent with Oryctolagus cuniculus、Ovis aries、Homo sapiens CCR10.【Conclusion】The Oryctolagus cuniculus CCR10 gene was successfully cloned in present study and the sequence has been submitted to GenBank(Accession. EU348829).