关键词: 百合, 百合无症病毒, 16 kD, 基因克隆, 原核表达, 抗血清制备

Abstract: Unknown gene（16 kD）was amplified by RT-PCR from Lily leaves infected by Lily symptomless virus and cloned into prokaryotic expression vector pET-28a（+）. Then the recombinant plasmid vector ligated with His-tag and carried 16 kD gene was transformed into E. coli strain BL21（DE3）. Induced with IPTG，the protein was highly expression in E. coli and the molecular weight of the recombinant protein 16 kD was 20 kD. After purification with Ni2+-NTA affinity chromatography，polyclonal antibody 16 kD was raised in mouses. Western blot analysis showed that the antiserum reacted specially with 16 kD protein of LSV；ELISA and RT-PCR also confirmed that the antiserum reacted specially with lily leaves infected by LSV. Our results indicate that the interest protein expression was detected，the antiserum reacted specially with 16 kD protein and used for LSV rapid test，immunohistochemistry and functional study of 16 kD protein.

Key words: lily, Lily symptomless virus, 16 kD, gene clone, prokaryotic expression, antiserum preparation