一个糖尿病肾病相关新基因的筛选、克隆和序列分析

Screening, Cloning and Analyzing of a New Diabetic Nephropathy Related Gene From db/db Mice

利用Affymetrix寡核苷酸基因表达谱芯片对 2型糖尿病肾病模型动物———db/db小鼠的肾脏基因表达谱进行了研究 .在此基础上 ,利用末端快速扩增法和RT PCR方法 ,对筛选出来的一个糖尿病肾病相关表达序列标签(EST)进行了cDNA克隆和表达分析 .得到了一长为 1 4kb的cDNA ,并暂时命名为mdnr4 1 1 (mousediabeticnephropathyrelatedmRNANo 4 1 1 ) .序列分析和网上数据库比对说明 ,这是一个新的cDNA序列 .利用DNA序列分析软件对此cDNA阅读框进行的预测性分析表明 ,此cDNA包含了一个完整的阅读框序列 .其编码的蛋白质由 90个氨基酸残基组成 .以氨基酸序列进行的同源性搜索表明 ,此多肽只与Archaeoglobusfulgidus的Na+ /H+ antiporter(napA 2 )具有局部的同源性 .以上结果表明 ,mdnr4 1 1是一个功能完全未知的小鼠糖尿病肾病相关新基因 .mdnr4 1 1cDNA的成功克隆 ,为进一步研究其生物学功能及其在糖尿病肾病发生。

Kidney gene expression profile of db/db diabetic neph ropathy mice was analyzed by using Affymetrix oligonucleotide genechip. Mice diabetic nephropathy related genes (and ESTs) were found out. Then, a 1 4 kb cDNA (mouse diabetic nephropathy related cDNA No 411, mdnr411, GenBank accession number AY256858) was amplified and cloned by using RACE method, based on the sequence of EST. Homologous analysis of the sequence was carried out. And the result shows that it is a new cDNA, which is highly homologous with mouse 'RIKEN cDNA 5830436L09 gene (5830436L09Rik) mRNA'. The ORF of mdnr411 was predicted by using DNASIS v 2 5 Demo program. The result shows that the peptide coded by mdnr411 mRNA has 90 amino acids. And the result of homology screening did not find any obviously homologous protein, except Archaeoglobus fulgidus Na+/H+ antipor ter (napA 2), which is partially homologous with the peptide coded by mdnr411 mRNA. All these results show that a cDNA of a new DNA related gene which function is fully unknown have screened and cloned. The foundation for further study of the function of the new gene, and its possible role in the pathogenesis of mice diabetic nephropathy has been laid, which is valuable to further understand the mechanism of diabetic nephropathy.