小鼠神经细胞p35^(nck5a)基因5′非编码区基因表达调控功能初步研究

Function of 5′flanking region of mouse neuronal cdc2-like kinase regulatory subunit p35^( nck5agene)

目的 :观察阿尔茨海默病 (AD)的相关基因 p35 nck5a基因 5′侧翼区指导报告基因表达的能力。方法 :将对本实验室所克隆的 p35 nck5a基因 5′侧翼片段连接于报告基因 β-半乳糖苷酶基因的上游构建表达载体 ,利用体外培养细胞和转基因小鼠实验体系来观察该片段在体内、外基因表达调控的差异。结果 :体外培养细胞中 p35 nck5a基因 5′侧翼序列不具有决定报告基因神经特异性表达的能力 ,但在基因组中整合有相同表达载体的转基因小鼠体内却能够指导报告基因的神经限制性表达。 结论 :这种差异提示进行体内。

Objective:To study the function of5′flanking region of mouse neuronal cdc2 - like kinase regulatory subunit p35 nck5agene. Methods:β- gal expression vector containing a part of 5′ flanking sequence(- 5 4 5 5 / - 90 ) was constructed. Transient expression assay in prim ary cerebrum cortex cell of rat (Neuron) and non- neuron cell,such as He L a,and transgenic mice were used to show the difference between in vivo and in vitro.Results:The fragm ent directed the expression ofβ- gal in Neuron and He L a cell,but in transgenic mice,expression ofβ- gal was found only in neural system.Conclusion:It is necessary to use experim ent system in vivo and in vitro to study the m olecular mechanism controling tissue- specific gene expression. [