【摘要】 [目的]克隆并分析广西巴马小型猪脂联素受体1(AdipoR1)和脂联素受体2(AdipoR2)编码基因的cDNA全序列。[方法]利用RT-PCR法,以骨骼肌总RNA为模板,分别扩增AdipoR1和AdipoR2全长cDNA序列,纯化后连接pMD 18-T载体,转化大肠杆菌DH5α,筛选阳性克隆体,鉴定后测序,并与其他物种AdipoR1和AdipoR2 cDNA序列进行同源性比较。[结果]RT-PCR扩增得到大小与AdipoR1和AdipoR2基因编码序列大小相同的片段,片段长度为1 128和1 161 bp。同源性比较分析发现,广西巴马小型猪AdipoR1和AdipoR2cDNA序列与GenBank中已报道的家猪脂联素受体同源性分别高达99.8%、99.7%,分别有1处和3处发生了碱基错义突变。[结论]该试验成功克隆了广西巴马小型猪AdipoR1和AdipoR2基因的cDNA,为进一步研究AdipoR基因的生物功能及设计以AdipoR为靶标的新型药物奠定了基础。更多还原

【Abstract】 [Objective] To clone and analyze the sequence of Adiponectin receptor 1(AdipoR 1) and receptor 2(AdipoR 2) cDNA of Guangxi Bama mini-pig.[Method] The Adiponectin receptors cDNAs were amplified by RT-PCR using skeletal muscle total RNA as template and then ligated into pMD18-T vector after purification.The recombinant pMD18-T vector was transformed into the E.coli DH5α for identification and sequencing.And the results were compared with the cDNA sequence from other species.[Result] The fragments,1 128 and 1 161 bp in size,were amplified by RT-PCR and respectively consistent with the coding sequence of AdipoR1 gene and AdipoR2 gene.The homology analysis showed that the sequences of AdipoR1 gene and AdipoR2 gene were respectively 99.8% and 99.7% homologous to the sequence of domestic pig reported in GenBank with 1 and 3 base missense mutations correspondingly.[Conclusion] The AdipoR1 gene and AdipoR2 gene were successfully amplified from Guangxi Bama mini-pig,laying the foundation for the further study of the biological function of AdipoR genes and the design of novel drugs with AdipoR genes as target.更多还原