CAJ | 학술논문

　　着-COP蛋白是真核生物分泌途径中COP玉有被小泡的一个亚基。本研究利用PCR技术扩增水稻着-c op基因(Os着-c op 1)的ORF(开放阅读框)，并克隆到原核表达载体pET-23d上，将表达载体pET-Os着-cop1转入大肠杆菌BL21(DE3)，以1.0 mmol/L的IPTG (isopropylβ-D-thiogalactoside)诱导表达重组蛋白，然后以重组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35 kD的重组蛋白，Western blot检测表明，免疫家兔的抗血清与水稻幼穗总蛋白杂交信号较好。Os着-COP1抗体的制备有助于研究该基因及COPI小泡在水稻中的功能。
　　착-COP단백시진핵생물분비도경중COP옥유피소포적일개아기。본연구이용PCR기술확증수도착-c op기인(Os착-c op 1)적ORF(개방열독광)，병극륭도원핵표체재체pET-23d상，장표체재체pET-Os착-cop1전입대장간균BL21(DE3)，이1.0 mmol/L적IPTG (isopropylβ-D-thiogalactoside)유도표체중조단백，연후이중조단백작위항원면역가토，제비다극륭항체。SDS-PAGE전영분석결과표명，성공유도표체료분자량약위35 kD적중조단백，Western blot검측표명，면역가토적항혈청여수도유수총단백잡교신호교호。Os착-COP1항체적제비유조우연구해기인급COPI소포재수도중적공능。
Theε-COP is one of the subunits of COPⅠvesicle in secret pathway of eukaryotic cells. The ORF (open reading frame) ofε-cop gene in rice (Osε-cop1) was obtained by PCR and cloned into the prokaryotic expres-sion vector pET-23d. The resulting plasmid was then introduced into E. coli strain BL21 (DE3) and induced to ex-press the recombinant Osε-COP1 protein by 1.0 mmol/L isopropyl?-D-thiogalactoside (IPTG). Afterwards, the re-combinant Osε-COP1 protein was used as antigen to immune rabbits. The results demonstrated a 35 kD protein was expressed successfully inducing by IPTG. Western blot analysis showed that the antiserum immunological recog-nized rice Osε-COP1 protein with high specificity, indicating that the polyclonal antibody was prepared successful-ly. This work would contribute to study the function of Osε-cop1 gene and COPⅠvesicle in rice.