中文摘要：

设计含有attB侧翼序列的引物，用于番木瓜环斑病毒（Papaya ringspot virus）的外壳蛋白基因（CP）基因的PCR扩增。在BP重组酶的作用下，将CP基因与受体载体pDONR^TM221发生重组反应，产生入门克隆。在LR重组酶作用下，将入门克隆与目的载体pHellsgate12发生重组反应，再进行平板培养筛选及PCR扩增和酶切反应。结果证明，用限制性内切酶Xho Ⅰ酶切消化后，琼脂糖凝胶电泳获得了预期长度为1700bp的重组片段；以启动子和外壳蛋白基因及外壳蛋白基因和终止子序列为基础，分别设计2对引物，经PCR扩增分别获得582和773bp片段。结果表明，获得了含有内含子的CP基因反向重复结构的表达载体。

英文摘要：

attB-flanked PCR primers were designed and used to amplify coat protein （CP） gene of Papaya ringspot virus by PCR. An entry clone was performed by a BP recombination reaction with attB-PCR products and donor vector pDONR^TM221, an expression clone was processed for plating selected with an LR recombination reaction with the entry clone and destination vector pHellsgate12 and ihpRNA （intron-containing hairpin） expression vector containing CP gene was testified by restriction enzyme digestion reaction and PCR. Results showed that the expected 1 700 bp recombination fragment was obtained by Xho Ⅰ restriction enzyme digestion of expression vector; the two pairs of primers were designed according to the sequence of 35S promoter and CP gene, and CP gene and OCS terminor, respectively, and two fragments（582 and 773 bp） were amplified by PCR. It indicated that IR expression vector of CP gene is constructed successfully.