Abstract In this experiment, we designed attB-flanked PCR primers and amplified coat protein gene (CP) and movement protein gene (CI) genes of Papaya ringspot virus with PCR, performed a BP recombination reaction with attB-PCR products and donor vector pDONRTM221 to generate an entry clone, then, processed an LR recombination reaction with the entry clone and destination vector pHellsgate12 of select to generate an expression clone system by screening of plate medium, and detected with PCR and enzyme digestion.