人脂联素基因的克隆及序列分析

Cloning and sequencing of human apM1 gene

目的:克隆人脂联素基因(apM1),为进一步研究脂联素的功能提供实验基础。方法:应用RT-PCR法自人大网膜脂肪组织的总RNA中扩增出apM1cDNA全长基因,采用T-A克隆法重组到pGEM-TVectorSystem中,通过蓝白斑筛选出阳性克隆后,测序鉴定。结果:从脂肪组织总RNA中扩增得到735bpapM1基因,其cDNA序列与GenBank报导的人脂联素基因apM1同源性为99.7%(159位和558位碱基发生了无义突变),获得了质粒pGEM-T-apM1,测序鉴定正确。结论:成功的克隆了apM1基因全长cDNA。

Objective： To clone human apM1 gene for the study molecular mechanism of the function of adiponectin. Methods： RT-PCRmethod was used to isolate, from the total RNA of human adipose tissue, cDNA fragment encoding adiponectin, which was linked intopGEM-T Vector, then amplified, and cloned. The sequence was analyzed to identify the recombinant plasmids. Resu/ts： The result of sequencing indicated that the sequence of the cloned cDNA was identical to that of the corresponding parts of the mRNA for human apM1 inGeneBank except 2 nucleotides. Conclusion： Human apM1 cDNA gene was successfully cloned.