【作者】 费菲； 王克双； 吕睦廸； 李捷； 程玉琴；

【Author】 Fei fei,Wang keshuang,L(u|¨) mudi,Li qie,Cheng yuqin (Pomology/Lab of Stress Physiology and Molecular Biology for Tree Fruits,A key Lab of Beijing Municipality,College of Agronomy and Biotechnology,China Agricultural University,Beijing 100193)

【机构】 中国农业大学农学与生物技术学院果树系/北京市果树逆境生理与分子生物学实验室；

【摘要】 利用特异性引物扩增得到了葡萄卷叶伴随病毒3号(Grapevine leafroll-associated virus 3,GLRaV-3)的外壳蛋白(coatprotein,CP)基因,并将其定向克隆到原核表达载体pET-28a(+)上,转化大肠杆菌BL21(DE3)。用终浓度为1mmol/L的IPTG进行诱导表达,可表达出分子量约为40 kDa的蛋白。将纯化的表达产物免疫家兔制备抗血清。通过对田间葡萄植株样品的ELISA和Western blot检测,证实了所制备的抗血清与GLRaV-3具有良好的特异性反应。更多还原

【Abstract】 The complete coat protein(CP) gene of Grapevine leafroll-associated virus 3 (GLRaV-3 )was amplified using specific primers,and then the gene was inserted into the vector pET-28a(+).The objective protein with the molecular weight of 40 kDa was expressed in E.coli BL-21(DE3) induced by IPTG at final concentration of 1 mmol /L.Antiserum was prepared after the rabbits were immunized with the purified recombinant CP.Indirect ELISA and Western blot confirmed that the antiserum reacted strongly and specially with CP of GLRaV-3 in field samples.更多还原