巴西橡胶树乳管细胞HbSKP1基因克隆及原核表达

Cloning and Prokaryotic Expression of HbSKP1 Gene from Latex in Hevea brasiliensis

SKP1蛋白是茉莉酸信号途径关键环节之一的SCFCOI1复合体的成员,在CUL1和COI1之间起连接作用。本文采用RT-PCR和RACE技术,从巴西橡胶树无性系热研7-33-97的胶乳中克隆了HbSKP1基因。该基因全长778bp,含一个543bp的开放阅读框,编码180个氨基酸。生物信息学分析表明,推导的HbSKP1氨基酸序列含有SKP1家族特征性的Skp1-POZ和Skp1结构域。HbSKP1与蓖麻、毛果杨、拟南芥、油棕、啤酒花、苜蓿的SKP1类蛋白的氨基酸序列同源性分别为64%、61%、47%、47%、47%和47%。原核表达了HbSKP1-His(6)融合蛋白。本研究为进一步鉴定乳管细胞中茉莉酸途径核心环节SCFCOI1复合体打下良好基础。

SKP1 protein is a component of SCFCOI1 complex and links CUL1 to COI1.HbSKP1 was cloned form rubber tree clone RY-7-33-97 by RT-PCR and RACE.The HbSKP1 contained a 543 bp open reading frame encoding a putative protein of 180 amino acids,HbSKP1.Two characteristic domains of SKP-like family,Skp1-POZ and Skp1 domains were present in the deduced amino acid sequence of HbSKP1.And furthermore,HbSKP1 shared 64%,61%,47%,47%,47% and 47% amino acid sequence identity with SKP-like proteins from Ricinus communis,Populus trichocarpa,Arabidopsis,Elaeis guineensis Jacg,Humulus lupulus and Medicago sativa Linn,respectively.The fused HbSKP1-His（6） was expressed in E.coli BL21 （DE3）.The results provide a good foundation for characterizing the compontents of SCFCOI1 complex,a key linker of JA signaling pathways in laticifer cells.