利用RT-PCR和RACE技术从蝴蝶兰中克隆的多酚氧化酶同源基因,命名为PhPPO,其cDNA全长1851bp,完整的编码框长1647bp,编码549个氨基酸,生物信息学分析表明其具有酪氨酸酶家族的特征,具有保守的CuA和CuB结合区,以及向叶绿体运输的导肽结构。将其亚克隆到表达载体pPRo-EXHTa上,在BL21(DE3)进行表达,经ITPG诱导,获得蝴蝶兰PPO原核表达蛋白。 The homologous gene of polyphenol oxidase cloned from Phalaenopsis by RT-PCR and RACE was named PhPPO. The full-length cDNA was 1851 bp in length and contained a complete coding sequence of 1647 bp, coding for 549 amino acids. Bioinformatics analysis showed that it had The tyrosinase family is characterized by the conserved CuA and CuB binding domains, as well as the peptide structure for transport to the chloroplast. Subcloned into expression vector pPRo-EXHTa, expressed in BL21 (DE3), and induced by ITPG to obtain the prokaryotic expression product of the phalaenopsis PPO.