草莓psy、pds和zds基因融合植物表达载体的构建  

Construction of plant binary expression vector of psy,pds and zds genes from strawberry

作　　者：朱海生[1,2,3] 陈敏氡[4] 温庆放[1,2,3] 

机构地区：[1]福建省农业科学院作物研究所 [2]福建省农业科学院蔬菜研究中心,福建福州350013 [3]福建省蔬菜工程技术研究中心,福建福州350013 [4]福建农林大学园艺学院,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2013年第4期391-397,共7页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金项目(31101534);福建省自然科学基金项目(2010J0111);福建省农业科学院科技创新团队重点科研项目(CXTD2011-20)资助

摘　　要：根据草莓psy、pds和zds基因序列,设计含有酶切位点的特异引物,以草莓总RNA为模板,通过RT-PCR法分别克隆psy、pds和zds基因.以植物表达载体pBI121为基础,利用口蹄疫病毒2A序列将psy、pds和zds基因融合在同一个开放阅读框下,构建成三价融合植物双元表达载体pBI2A2Apsy-pds-zds.同时构建了psy和zds基因二价融合植物表达载体pBI2A2Apsy-zds.经PCR、限制性内切酶酶切和测序鉴定后,成功将2个重组表达质粒pBI2A2Apsy-pds-zds和pBI2A2Apsy-zds导入农杆菌EHA105中.Specific primers containing different enzyme sites were designed on the basis ofpsy, pds and zds genes sequence of straw- berry. With the template of total RNA, psy, pals and zds genes were amplified by RT-PCR. Based on the vector of pBI121, the tri- valent plant express vector pBI2A2Apsy-pds-zds containing psy, pds and zds genes and fused in a single open reading frame （ORF） by foot and mouth disease virus （FMDV） was constructed. In addition, the divalent expression vector pBI2A2Apsy-zds containing psy and zds genes was also constructed. Vectors of pBI2A2Apsy-pds-zds and pBI2A2Apsy-zd, s were checked by PCR, restriction en- zymes analysis and sequencing, and transformed into Agrobactrium tumefaciens EHA 105.

关 键 词：草莓 psy PDS zds 表达载体 

分 类 号：S668.4[农业科学—果树学]