FAD2和γ-TMT基因双价种子特异表达载体的构建及对花生的转化

Seed-specific expression vector of FAD2 and γ-TMT gene construction and transformation in peanut

根据已发表的γ-TMT基因序列(GenBank AF104220),从拟南芥(Arabidopsis thaliana)的cDNA中分离得到γ-TMT基因,根据已发表的油菜(Brassica napus)种子贮藏蛋白基因特异表达的2S启动子(登录号J02798)序列,从油菜DNA中分离得到2S启动子。为了同时提高花生中维生素E和油酸含量,构建得到了双价种子特异表达载体pCAMBIA1300AT,它带有FAD2基因反义RNA干扰体和γ-TMT基因的种子特异表达单元。该载体已登录在GenBank上(登录号FJ362601)。通过农杆菌C58介导转化花生品种闽花6号,获得PCR阳性的转化植株。

A plant expressing binary vector pCAMBIA1300AT[GenBank accession number FJ362601]was constructed containing seed specific 2S promoter controlled γ-TMT gene and an antisense RNA expressing element FAD2. It was used to increase both tocopherol and oleic acid content in peanut through Agrobacterium tumefaciens mediated transformation. In the vector,γ-TMT gene was isolated from Arabidopsis thaliana cDNA according to GenBank sequence AF104220. 2S promoter was isolated from Brassica napus DNA according to J02798 sequence, which controls a seed storage protein. After transformation,putative transformants of peanut cultivar Minghua 6 were obtained after PCR identification.