高温致金黄地鼠神经管畸形的上调表达基因筛选及其鉴定  

Cloning and characterization of up-regulated genes in golden hamsters with neural tube defects induced by hyperthermia

作　　者：杨青[1] 高英茂[1] 李少玲[1] 

机构地区：[1]山东大学医学院组织胚胎学教研室,济南250012

出　　处：《解剖学杂志》2007年第2期196-199,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金(30270701)

摘　　要：目的:筛选高温致金黄地鼠神经管畸形的上调表达基因,探讨其分子生物学机制。方法:通过抑制性消减杂交技术获得消减产物,转化大肠杆菌DH5α,构建高温致金黄地鼠胚胎神经管畸形的消减cDNA文库;联合蓝白斑和菌落PCR方法筛选含插入片段的阳性克隆,进行测序和同源性分析,并经Northern杂交技术检测基因的表达。结果:成功构建高温致金黄地鼠胚胎神经管畸形的消减cDNA文库,其中2个克隆插入片段的基因序列与小鼠磷酸甘油酸酯激酶(Pgk-1)同源。Northern杂交证实该基因在高温致畸胚胎神经管的表达较其在同龄正常胚胎神经管明显升高。结论:Pgk-1的上调表达可能与高温致神经管畸形密切相关。Objective： To clone up-regulated genes in the golden hamster with neural tube defects induced by hyperthermia and explore the related molecular mechanism. Methods： Subtracted eDNAs was obtained using suppression subtractive hybridization（SSH）. After transformation of the subtracted eDNAs into E eoli, DH5α, a subtraetive library was constructed. Clones with inserts were selected with combination of α-eomplementary phenomenon and colony PCR method. The sequencing and homology analysis were then performed and the mRNA expression levels were measured by Northern blotting. Results： A subtraetive library was successfully constructed, including two eDNA clones with inserts homologous to mouse phosphoglyeerate kinase 1（Pgk-1）. Northern blotting demonstrated that Pgk-1 was expressed higher in the defected neural tubes induced by hyperthermia than that in the normal developed ones. Conclusion： The higher expression level of Pgk-1 may be strongly related to neural tube that defects induced by hyperthermia in golden hamsters.

关 键 词：神经管畸形 高温 抑制性消减杂交 基因 金黄地鼠 

分 类 号：R363[医药卫生—病理学]