水稻 OsCER4 基因编码脂肪醛脱羧酶，与拟南芥 CER1基因高度同源，是否参与植物表面蜡质合成尚不清楚,为了探讨其功能，扩增了 OsCER4 基因起始密码子ATG上游约2 kb的序列作为该基因的启动子，以常规技术将启动子和 OsCER4 基因反义片段克隆到pCAMBIA双元载体1380中，采用农杆菌介导法转化粳稻品种中花11，并进行了转化苗 OsCER4 蛋白的表达量变化分析.结果表明，成功构建了由 OsCER4 自身启动子驱动的反义RNA载体，并通过农杆菌介导法成功转入水稻中花11中，多数 OsCER4 基因反义转化植株为阳性转化植株，后代分离比符合 3∶1，反义RNA转化植株在蛋白水平上的表达量下降.
OsCER4 gene is a member of fatty aldehyde decarbonylase gene family and is highly homologous to CER1 gene in Arabidopsis, however, it is still not known whether OsCER4 participates in the synthesis of cuticular wax. A 1946-bp genomic fragment located on the upstream of the annotated ATG start codon was amplified and used as cognate promoter for antisense RNA vector construction, and a 624-bp fragment of the OsCER4 cDNA was amplified from total RNAs isolated from Zhonghua 11, and then the cognate promoter and OsCER4 cDNA fragments were cloned into the pCAMBIA1380 which was transformed into rice by Agrobacterium-mediated method. The results showed that the antisense transgene driven by the cognate promoter was constructed and transformed into rice successfully, and most of the OsCER4 antisense RNA transgenic plants were positive and segregated as single T-DNA insertion by PCR and segregation analysis of Hyg-resistant in T1 progeny, and the expression of OsCER4 significantly decreased in the OsCER4 antisense RNA transgenic plants by Western blotting.
周玲艳,姜大刚,李 静,周 海,庄楚雄.水稻蜡质合成相关基因 OsCER4 自身启动子驱动的反义RNA转化植株获得[J].华南农业大学学报,2013,34(1):51-56复制