无花果ACS1基因RNAi植物表达载体构建  

RNA interference expression vector construction of Ficus carica ACS1

作　　者：唐霞[1] 张帅[1] 马俊莲[1] 张子德[1] 

机构地区：[1]河北农业大学食品科技学院,河北保定071001

出　　处：《河北农业大学学报》2013年第4期62-65,共4页Journal of Hebei Agricultural University

摘　　要：通过构建无花果ACS1基因的RNA干扰植物表达载体,为研究ACS基因的功能及采用生物技术方法培育无花果耐贮新品种奠定基础。本试验根据已克隆的无花果ACS1基因序列及植物表达载体pBI221、pBI121的酶切位点,设计2对带相应酶位点的特异性引物,以测序质粒为模板,进行PCR扩增,获得正、反向扩增片段,分别正、反向插入到pBI221载体中的CaMV35S启动子和gusA基因之间,构建成无花果ACS1基因的RNAi中间表达载体。再将中间载体中的正、反向片段和gusA基因以双酶切方式连接到表达载体pBI121上,构建成pBI121-RNAi-ACS1植物表达载体。通过各种限制性内切酶的酶切鉴定,成功构建了无花果ACS1基因的RNA干扰植物表达载体。The RNA interference expression vector construction of Ficus carica ACS1 gene is beneficial to exploring the function of the gene and the biological technology to improve the breeding method of Ficus carica cultivars. According to the restriction enzyme sites of expres- sion vector and cloning sequence of Ficus carica ACS1 gene, two pairs of primers were de- signed which contain restriction enzyme sites and used to amplify sequenced plasmid, and then the sense and antisense PCR products were obtained. The PCR products were inserted between the CaMV 35S promoter and gusA gene into the expression vector pBI221. The three gene fragments of RNAi structure including the sense and antisense fragments of ACS1 gene and gu- sA gene were digested by restricted enzymes, then linked into the expression vector pBI121. The RNAi expression vector called pBI121-RNAi-ACS1 was obtained. Through restriction en- zyme identification,the RNAi plant expression vector of Ficus carica ACS1 gene was success- fully constructed.

关 键 词：无花果 ACS1基因 植物表达 

分 类 号：S663.3[农业科学—果树学]