甘蓝型油菜TT1基因反义植物表达载体的构建  

Construction of A Plant Expressing Vector for Antisense Suppression of TT1 Gene Family in Oilseed Rape (Brassica napus)

作　　者：路小春[1] 李加纳[1] 李本逊[2] 柴友荣[1] 

机构地区：[1]西南大学农学与生命科学学院农业部生物技术与作物品质改良重点实验室 [2]云南农业大学农学与生物技术学院,昆明650201

出　　处：《生物技术通报》2006年第C00期305-309,共5页Biotechnology Bulletin

基　　金：国家自然科学基金重点项目(30330400);重庆市自然科学基金重点项目(8446)资助

摘　　要：拟南芥TT1(transparenttesta1)基因编码一个WIP类型锌指蛋白转录因子,调控内种皮发育和原花青素在内种皮的积累。本研究通过PCR扩增了甘蓝型油菜(Brassicanapus)TT1基因家族(BnTT1)的一段特异保守片段TT1A,并将其亚克隆到中间载体pCambia2301G中,构建了TT1反义植物表达载体pCambia2301G-TT1A,通过PCR鉴定、酶切鉴定和DNA测序证实载体构建成功,并转化到根癌农杆菌LBA4404菌株中形成工程菌株。此表达载体的构建为进一步研究TT1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利用反义TT1基因遗传转化获得转基因新型黄籽油菜奠定了基础。TT1 （ transparent testa 1 ） gene of Arabidopsis thaliana encodes a WIP-type zinc finger transcription factor. It regulates endothelium development and proanthocyanidin accumulation in the seed coat. A fragment TT1A corresponding to a specific conserved region of TT1 gene family of Brassica napus （ BnTT1 ） was amplified by PCR and sub- cloned into intermediate vector pCambia2301G to form plant expression vector pCambia2301G-TT1A, as 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 TT1 genes involved in yellow seed trait formation, as well as for creating novel yellow seed germplasm via suppressed TT1 expression in B. napus.

关 键 词：TT1 类黄酮 反义 载体 甘蓝型油菜 

分 类 号：S565.4[农业科学—作物学]