大鼠视觉相关脑区表达序列标签的克隆与鉴定

Cloning and Identification of E S T from Vision Associated Region of Rat Brain

目的 分析视觉相关脑区基因的差异表达并获取新的表达序列标签。方法 利用差异显示反转录 Ｐ Ｃ Ｒ（ Ｄ Ｄ Ｒ Ｔ Ｐ Ｃ Ｒ）技术分析大鼠外侧膝状体与视皮层基因差异表达情况，并筛选差异表达基因片段。结果 经 Ｄ Ｄ Ｒ Ｔ Ｐ Ｃ Ｒ筛选并通过反 Ｎｏｒｔｈｅｒｎ 杂交去除假阳性，最终获得 １ 个差异表达基因片段。对其进行克隆、测序和同源性比较发现，该片段为一新的表达序列标签，并与心肌细胞集钙蛋白的 ｍ Ｒ Ｎ Ａ 具有 ７４％ 的同源性。结论 视觉相关脑区差异表达序列的分析可能为研究视觉形成的分子机制或视觉通路的分子构筑提供有益的线索。

Objective To analyze the differential gene expressions from vision associated region of rat brainand to screen a new expressed sequence tag ( E S T) . Methods Differential display reverse transcriptionpolym erase chain reaction ( D D R T P C R) w as carried out to screen the differentially expressed gene frag m ents from the lateralgeniculate body and the visual cortex. Results One differentially expressed fragm entw as obtained and proved to be true positive through reverse northern assay. After cloning, sequencing andhom ology com paring, the fragm ent w as found to be a novel E S T and show ed 73% hom ology w ith cardiaccalsequestrin m R N A . Conclusion The results provide a candidate gene and the function of w hich m ay be aclue for illum inating m olecular m echanism s of vision or m olecular construction of visualpathw ay.