甘蓝型油菜F3’H基因反义植物表达载体的构建

类黄酮3’-羟化酶(F3’H)是类黄酮途径中的一个关键酶,对种皮色泽和花色的形成具有重要作用。通过PCR法扩增了甘蓝型油菜(Brassica napus L.)F3’H基因的一段特异保守片段F3’HA,并将其亚克隆到中间载体pCAMBIA2301G中,构建了F3’H反义植物表达载体pCAMBIA2301-F3’HA,通过PCR鉴定和酶切鉴定证实载体构建成功,并转化到根癌农杆菌LBA4404菌株中形成工程菌株。为进一步研究F3’H基因在甘蓝型油菜种皮色素合成途径中的分子生物学机理和该基因的代谢调控网络奠定基础。

Construction of An Antisense Plant Expression Vector of F3'H from Rapeseed (Brassica napus L.)

Flavonoid 3'-hydroxylase(F3'H)is a key enzyme of flavonoid pathway and play important roles in determining the color of testa and flowers.A specific conserved region fragment of F3'H from Brassica napus was amplified by PCR and subcloned into intermediate vector pCAMBIA2301G to form an antisense plant expression vector pCAMBIA 2301G-F3'H,proved by PCR analysis,endonuclease digestion and sequencing.Then the recombination vector was introduced into Agrobacterium tumefaciens strain LBA4404 to form an engineering strain.This work lays a basis for clarifying the molecular mechanism of F3'H genes involved in yellow seed trait formation,as well as for elucidation of the formation of yellow seed trait and known of the function of the F3'H.