AC2 gene of Tomato yellow leaf curl Guangdong virus isolate G3 (TYLCGuV-[G3]) was amplified from its recombinant plasmid by PCR, and the gene was cloned into prokaryote expression vector pET-28b( + ). The expressed fusion protein was highly expressed in Escherichia coli Rosetta( DE3) Ⅲ induced by IPTG, and the molecular mass was about 19 500 by SDS-PAGE analysis. The fusion protein was used as an antigen to immunize the healthy rabbit and the antiserum of AC2 was prepared. The titer measured by ELISA was about 1: 16 384. Western-blotting analysis indicated that the antiserum could se-rologically react with AC2 protein of TYLCGuV-[ G3].%利用PCR方法从广东番茄黄化曲叶病毒分离物G3(Tomato yellow leaf curl Guang dong virus-[ G3],TYLCGuV-[G3])全长序列的重组质粒上扩增该病毒的AC2基因,将其构建至原核表达载体pET-28b(+)上,经IPTG诱导,在大肠埃希菌Rosetta( DE3)Ⅲ中高效表达.SDS-PAGE电泳结果显示,目的融合蛋白与预期大小一致,相对分子质量约为19 500.以诱导的融合蛋白为抗原,免疫注射新西兰大耳白兔制备抗血清,间接ELISA测定抗血清效价为1∶16 384.Western-blotting检测结果表明,制备的抗血清有较好的抗原-抗体识别反应,为专化性抗血清.
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