CAJ | 학술논문

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）B121（DE3）菌株，以1．2mmol／L的异丙基-β-D-硫代吡喃半乳糖苷（Isopropyl β-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71．6kD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．0sCER4抗体的制备有助于研究OsCER4基因的表达特性．
이용PCR방법확증수도（Oryza sativa L．）사질기인OsCER4적개방열독광（Open reading frame，ORF），병극륭도원핵표체재체pET-23d상，장표체재체OsCER4-pET전입대장간균（Escherichia coli）B121（DE3）균주，이1．2mmol／L적이병기-β-D-류대필남반유당감（Isopropyl β-D-thiogalactoside，IPTG）유도표체융합단백，연후이융합단백작위항원면역가토，제비다극륭항체．SDS-PAGE전영분석결과표명，성공유도표체료분자량약위71．6kD적융합단백，이Western blot검측표명，면역가토적항혈청여수도유묘총단백잡교신호교호．0sCER4항체적제비유조우연구OsCER4기인적표체특성．
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.