激光生物学报, 2011, 20 (6): 798, 网络出版: 2015-10-08  

人类新基因Bzw2的克隆、抗体制备及表达分析

Cloning, Polyclonal Antibody Preparation and Expression Analysis of Human Novel Gene Bzw2
作者单位
湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室, 心脏发育研究中心, 湖南 长沙410081
摘要
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GST-Bzw2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。
Abstract
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 Bzw2 in the process of heart development, by using the bioinformatics methods, the full length of Bzw2 gene was cloned and inserted into pGEXT-4T-1 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 Bzw2 polyclonal antibody was identified by Western blot. Western blot was investigated by using mouse tissues. The result shows that Bzw2 is highly expressed in mouse heart, muscle and brain.

姚力群, 李志, 江志钢, 万永奇, 吴秀山, 袁婺州. 人类新基因Bzw2的克隆、抗体制备及表达分析[J]. 激光生物学报, 2011, 20(6): 798. YAO Liqun, LI Zhi, JIANG Zhigang, WAN Yongqi, WU Xiushan, YUAN Wuzhou. Cloning, Polyclonal Antibody Preparation and Expression Analysis of Human Novel Gene Bzw2[J]. Acta Laser Biology Sinica, 2011, 20(6): 798.

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