单侧视觉剥夺后鸽脑两侧视盖基因的差异表达

Differential Expression of Gene in Optic Tecta of Pigeon after Monocular Deprivation

目的 探讨单侧视觉剥夺后鸽子两侧视盖基因的差异表达情况。方法 建立单眼剥夺动物模型 ,抽提、纯化左右视盖mRNA ,利用SSH方法建立左侧视盖差异表达文库 ,对其中EST进行克隆、测序 ,反Northern杂交去除假阳性。结果 共筛选得到 12个差异表达片段 ,经同源性检索 ,其中 11个未发现有高同源性的基因 ,提示可能为新的EST ;另一差异片段与CGI- 3 1蛋白有 85 %的同源性。

Objective To study the differential expression of the pigeon optic tecta gene after depriving its left eye.Methods A model of monocular deprivation of left eye was constructed, RNAs of right and left optic tecta were isolated, and mRNA was extracted. The subtractive library of left optic tecta was constructed by suppression subtractive hybridization(SSH). The expressed sequence tags (EST) were cloned and sequenced, while the false positive clones were eliminated by reverse Northern assay.Results 12 of 24 clones were identified to be true positive. Of these 12 clones, 11 were proved to be new cDNA fragment and 1 showed 85% homology with homo sapiens CGI 31 protein mRNA.Conclusion This results provide candidate genes which may be a clue for illuminating molecular mechanism of vision or molecular construction of vision pathway.