水稻蜡质基因OsCER4的原核表达及多克隆抗体制备  

Prokaryotic expression and polyclonal antibody preparation of wax gene OsCER4in rice

作　　者：周玲艳[1,2] 秦华明[3] 周海[2] 杨家伟[2] 庄楚雄[2] 

机构地区：[1]仲恺农业工程学院生命科学学院,广东广州510225 [2]华南农业大学生命科学学院,广东广州510642 [3]暨南大学理工学院,广东广州510564

出　　处：《仲恺农业工程学院学报》2012年第1期11-15,共5页Journal of Zhongkai University of Agriculture and Engineering

基　　金：广东省自然科学基金(S2011040001653)资助项目

摘　　要：利用PCR方法扩增水稻(Oryza sativa L.)蜡质基因OsCER4的开放阅读框(Open reading frame,ORF),并克隆到原核表达载体pET-23d上,将表达载体OsCER4-pET转入大肠杆菌(Escherichia coli)BL21(DE3)菌株,以1.2 mmol/L的异丙基-β-D-硫代吡喃半乳糖苷(Isopropylβ-D-thiogalactoside,IPTG)诱导表达融合蛋白,然后以融合蛋白作为抗原免疫家兔,制备多克隆抗体.SDS-PAGE电泳分析结果表明,成功诱导表达了分子量约为71.6 kD的融合蛋白,而Western blot检测表明,免疫家兔的抗血清与水稻幼苗总蛋白杂交信号较好.Os-CER4抗体的制备有助于研究OsCER4基因的表达特性.The ORF of OsCER4 gene in rice was obtained by PCR and cloned into the prokaryotic expres- sion vector pET-23d, then the resulting recombinant vector was introduced into Escherichia coli strain BL21 （DE3） and induced to express the OsCER4 fusion protein by 1.2 mmolfL isopropyl-β-D-thiogal- actoside （IPTG）. The OsCER4 fusion protein was used as antigen to immune rabbits. The results showed that the protein of 71.6 kD was expressed successfully by induction with IPTG and the polyclonal antibodies were obtained by using OsCER4 fusion protein as antigen to immune rabbits and the immuno- logical specificity of the polyclonal antibody to OsCER4 was confirmed by Western blot analysis. The preparation of OsCER4 antibodies was contributed to detect the expression pattern of OsCER4 gene in rice.

关 键 词：水稻(Oryza SATIVA L.) OsCER4基因 原核表达 多克隆抗体 

分 类 号：Q511[生物学—生物化学]