CAJ | 학술논문

通过RT－PCR方法扩增获得小麦黄花叶病毒（ Wheat yellow mosaic virus，WYMV）的衣壳蛋白（ CP）基因，将其连接原核表达载体pEHISTEV，并将重组质粒转化大肠杆菌Rosetta，经IPTG诱导后，可以表达38 kD的融合蛋白。通过切胶回收的方法收集目的蛋白，免疫新西兰长耳兔，制备WYMV CP的多克隆抗体。间接ELISA测定该抗血清效价为1∶2048，Western blotting分析表明该血清可以与病株中CP发生特异性反应，而与健康植株汁液无反应。该血清为WYMV的检测及CP功能的研究奠定了基础。
통과RT－PCR방법확증획득소맥황화협병독（ Wheat yellow mosaic virus，WYMV）적의각단백（ CP）기인，장기련접원핵표체재체pEHISTEV，병장중조질립전화대장간균Rosetta，경IPTG유도후，가이표체38 kD적융합단백。통과절효회수적방법수집목적단백，면역신서란장이토，제비WYMV CP적다극륭항체。간접ELISA측정해항혈청효개위1∶2048，Western blotting분석표명해혈청가이여병주중CP발생특이성반응，이여건강식주즙액무반응。해혈청위WYMV적검측급CP공능적연구전정료기출。
The coat protein ( CP) gene of wheat yellow mosaic virus ( WYMV) was amplified by RT-PCR and cloned into the prokaryotic expression vector pEHISTEV to produce recombinant plasmid pEHISTEV-WYMVCP.The recombinant plasmid was transformed into Escherichia coli strain Rosetta and expressed a 38 kD fusion protein after inducing by IPTG.The fusion protein was collected as antigen to immunize rabbits for production of polyclonal antiserum against WYMV CP.Finally the polyclonal antiserum was obtained with titer above 1∶2 048 by ELISA.With this antiserum, the WYMV CP in the infected wheat plant could be detected specifically by Western blotting.The resultant antibody will facilitate the detection of WYMV and the function-al studies of WYMV CP.