补锌糖尿病大鼠差异显示新基因片段的克隆鉴定及组织表达

Cloning,identification and tissue expression of novel genes differentially expressed in diabetic rats supplemented with zinc

目的克隆补锌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分布。方法对前期研究中分离到的与补锌有关的糖尿病大鼠差异显示的基因片段进行克隆、测序,同源性分析,对发现的2条新基因(2#差异显示片段及6#差异显示片段)设计特异性引物,进行RT-PCR检测,以观察新基因在各组大鼠肝脏中的表达变化,并检测新基因在不同组织中的表达情况。结果经克隆、测序、同源性分析,在GenBank中未找到与2#差异显示片段及6#差异显示片段高同源性的序列,确定2#及6#差异显示片段为新的基因片段;2#及6#新基因片段在糖尿病对照组、糖尿病补锌组的表达量均低于正常对照组,糖尿病补锌组的表达量高于糖尿病对照组(P<0.05);2#新基因片段在大脑及胰腺中表达量较高,在肾脏中表达量较低,在心肌、骨骼肌、胸腺中无表达。6#新基因片段在心肌、骨骼肌、大脑、肾脏、胰腺中均有表达,但在胸腺中无表达;2#及6#新基因片段被GenBank收录,接收号分别为AY952968、AY952970。结论分离并克隆了2条与补锌有关的糖尿病大鼠差异显示的新基因片段,经RT-PCR检测其在各组大鼠肝脏及其它各组织中表达分布后,在GenBank上成功登录。

Objective To clone novel gene fragments differentially expressed from diabetic rats supplemented with zinc and to detect their expression distribution in various tissues. Methods cDNA fragments from former research project were cloned, sequenced and BLASTn analysed. The RT-PCR of the two 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 2 # cDNA and 6 # cDNA were shown to be the novel gene fragments for no matched gene with them in GenBank. The expression level of DM group and DM ＋ Zn group was obviously lower than those of NC group. The expression level of DM ＋ Zn group was higher than those of DM group （ P 〈 0.05）. The expression level of 2 # cDNA was higher in brain and pancreas, lower in kidney, and none in heart, muscle and thymus. The expression of 6 # cDNA can be found in heart, muscle, brain, kidney and pancreas, but thymus. 2 # cDNA and 6 # cDNA were registered to GenBank with the numbers of AY952968 and AY952970 respectively. Conclusion Two novel genes differentially expressed in diabetic rats supplemented with zinc were cloned and expressed in various tissues, and were successfully submitted to GenBank.