甘蓝型油菜透明种皮12(TT12)基因家族反义抑制植物表达载体的构建  

Construction of A Plant Expressing Vector for Antisense Suppression of TRANSPARENT TESTA 12 (TT12) gene Family in Oilseed Rape (Brassica napus)

作　　者：黄华磊[1] 雷波[1] 柴友荣[1] 

机构地区：[1]西南大学农学与生物科技学院,农业部生物技术与作物品质改良重点实验室,重庆市作物品质改良重点实验室,重庆市油菜工程技术研究中心,重庆400716

出　　处：《中国农学通报》2007年第5期43-48,共6页Chinese Agricultural Science Bulletin

基　　金：国家863项目"甘蓝型油菜类黄酮途径功能基因克隆及黄籽性状代谢工程研究"(20060110Z1024);重庆市自然科学基金重点项目"甘蓝型黄籽油菜品质形成的分子生物学机理研究"(8446);国家自然科学基金重点项目"甘蓝型油菜优质高效黄籽性状的遗传基础研究"(30330400)

摘　　要：【研究目的】构建一个同时反义共抑制甘蓝型油菜(Brassica napus)透明种皮12基因家族(BnTT12)转录的植物表达载体。【方法】通过PCR扩增得到BnTT12家族一段503bp的特异保守片段TT12A,通过亚克隆整合到中间载体pCambia2301G,构建TT12反义植物表达载体pCambi-a2301G-TT12A。采用液氮冻融法将其转化到根癌农杆菌。【结果】经过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TT12反义表达载体的工程菌株。【结论】pCambia2301G-TT12A的成功构建为利用此反义载体通过遗传转化获得转基因新型黄籽油菜和进一步探明TT12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奠定了基础。[OBJECTIVE]To construct an antisense plant vector simultaneously suppressing the transcription of Brassica napus TRANSPARENT TESTA 12 （BnTT12） gene family.[METHOD]A fragment TT12A corresponding to a specific conserved region of BnTT12 gene family was amplified by PCR and subcloned into intermediate vector pCambia2301G to form plant expression vector pCambia2301G-TT12A. [RESULTS]As proved by PCR analysis, endonuclease digestion and sequencing, the construction of pCambia2301G-TT12A was successful. The recombination vector was introduced into Agrobacterium tumefaciens strain LBA4404 to form an engineering strain. [CONCLUSION]pCambia2301G-TT12A lays a basis for creating novel yellow seed germplasm via suppressed TT12 expression, as well as for clarifying the molecular mechanism of TT12 genes involved in yellow seed trait formation in B napus.

关 键 词：TT12 类黄酮 反义 载体 甘蓝型油菜 

分 类 号：Q16[生物学—普通生物学]