人类新基因Bzw2的克隆、抗体制备及表达分析

摘要

Bzw2 ( basic leucine zipper and W2 domains 2 ) gene is a candidate gene for heart development. It contains bzip domain and W2 domain. To further investigate the possible functions of Bzwl in the process of heart development, by using the bioinformatics methods, the full length of Bzwl gene was cloned and inserted into Pgext-4T-l vector. The GST-Bzw2 fusion protein was induced by IPTG. After purification, the fusion protein was used to immunize the New Zealand white rabbits. The titer of Bzwl polyclonal antibody was identified by Western blot. Western blot was investigated by using mouse tissues. The result shows that Bzwl is highly expressed in mouse heart, muscle and brain.%Bzw2( basic leucine zipper and W2 domains 2)基因是人类心脏发育候选基因,它由bzip结构域和W2结构域构成.为了研究该基因在心脏发育过程中的作用,利用生物信息学信息克隆了人类Bzw2基因全长,将所得的片段插入到原核表达载体pGEXT-4T-1载体中,利用BL21菌株表达该重组质粒,经过IPTG诱导,得到GSTBzw2融合蛋白,蛋白经纯化分离后免疫新西兰大白兔,并用Western blotting检测抗体效价.利用抗体进行了心脏,肌肉,脑组织表达检测,结果显示,Bzw2在心脏,肌肉,脑组织中高表达.