Abstract: 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.

Key words: Arachis hypogaea, AhAO1, pPROEXHTa, prokaryotic expression, antibody

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。

关键词: 花生, AhAO1, pPROEXHTa, 原核表达, 抗体