补硒糖尿病大鼠新基因片段的鉴定及组织表达

IDENTIFICATION AND TISSUE EXPRESSION OF NOVEL GENES IN DIABETIC RATS SUPPLEMENTED WITH SELENIUM

目的:克隆补硒糖尿病大鼠差异显示的新基因片段并检测其在各组织中的表达水平。方法:对前期研究中分离到的补硒糖尿病大鼠与糖尿病对照组之间差异显示的基因片段进行克隆、测序、同源性分析,对发现的5条新基因(Se-2、Se-6、Se-10、Se-14、Se-18)设计特异性引物,进行RT-PCR检测,以排除假阳性并观察其在各组大鼠肝脏中的表达水平及在不同组织中的表达分布。结果:经克隆、测序、同源性分析,发现有5个基因片段(Se-2、Se-6、Se-10、Se-14、Se-18)在GenBank中未找到高同源性的序列,为新的EST片段。其中4个基因片段(Se-2、Se-10、Se-14、Se-18)在糖尿病对照组、糖尿病补硒组肝脏中的表达量均低于正常对照组,糖尿病补硒组的表达量高于糖尿病对照组(P<0.05);在大鼠多种组织中表达水平也不甚相同。4个新基因片段被GenBank收录,接收号分别为AY952968、DQ351839、DQ351838、DQ351840。结论:分离并克隆的4条与补硒有关的糖尿病大鼠差异显示的新基因片段,可能与硒改善糖尿病糖脂代谢紊乱有一定关系。

Objective： To clone novel gene fragments differentially expressed from diabetic rats supplemented with selenium and detect their expression distribution in various tissues. Method： cDNA fragments from former research project were cloned, sequenced and BLASTn analysed. The RT-PCR of the five novel genes were made using the primers designed according to the sequence of cDNA to observe the expression changes in liver of various groups and their expression distributions in various tissues. Results： Se-2, Se-6, Se-10, Se-14 and Se-18 cDNA were shown to be the novel gene fragments for no matched gene with them in GenBank. The expression levels of four cDNAs, including Se-2, Se-10, Se-14, Se-18, in DM group and DM＋Se group were obviously lower than those in NC group. The expression level of DM＋Se group was higher than those in DM group （P〈0.05）. The expression levels of the novel cDNAs were different in heart, muscle, brain, kidney, pancreas, and thymus. They were registered to GenBank with the numbers of AY952968, DQ351839, DQ351838 , DQ351840, respectively. Conclusion： The four novel genes differentially expressed in diabetic rats supplemented with selenium were cloned and expressed in various tissues, and were successfully submitted to GenBank.