花生AhAO1蛋白的原核表达、纯化及抗体制备  

Prokaryotic Expression,Purification and Polyclonal Antibody Preparation of Peanut AhAO1 Protein

作　　者：杨丽霞[1,2] 宋涛平[1] 彭新凯[1] 胡朝晖[1] 李玲[2] 

机构地区：[1]长沙市食品质量安全监督检测中心,湖南长沙410013 [2]华南师范大学生命科学学院广东省植物发育生物工程重点实验室,广东广州510631

出　　处：《亚热带植物科学》2010年第4期4-6,共3页Subtropical Plant Science

基　　金：教育部科学技术研究重点项目(03098)

摘　　要：采用RT-PCR方法,以花生叶片RNA逆转录得到的cDNA为模板,扩增出AhAO1基因片段,将其插入原核表达载体pPROEXHTa中,构建AhAO1基因片段原核表达载体HTaAhAO1,经PCR和测序确证后,以IPTG诱导其在E.coli BL21中高表达His-AhAO1融合蛋白,采用亲和层析柱与电泳纯化融合蛋白。用纯化的His-AhAO1融合蛋白制备抗体,为分析AhAO1的功能奠定基础。The AhAO1 gene was isolated by RT-PCR using cDNA synthesized from total RNA extracted from peanut leaves and cloned into plasmid pPROEXHTa.The expression plasmid HTaAhAO1 was transformed into E.coli BL21.SDS-PAGE analysis revealed that the His-AhAO1 fusion protein was highly expressed in E.coli BL21 and purified by affinity chromatography and electrophoresis.The His-AhAO1 polyclonal antibody was prepared and tested by western blot.The anti-His-AhAO1 polyclonal antibody lays foundation for further research on AhAO1.

关 键 词：花生 AhAO1 pPROEXHTa 原核表达 抗体 

分 类 号：Q943.2[生物学—植物学]