aFGF植物表达载体构建及转化紫花苜蓿的初步研究  

A rudimentary study of the acid fibroblast growth factor's plant expression vector construction and transformation into Medicago sativa

作　　者：蒋世翠[1] 王义[1] 孙春玉[1] 王艳芳[1,2] 李校堃[1,2] 张美萍[1] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]国家教育部生物反应器工程中心,吉林长春130118

出　　处：《草业学报》2011年第4期180-186,共7页Acta Prataculturae Sinica

基　　金：十一五"863"计划生物反应器重大专项(No.2007AA100503);吉林省科技发展重点计划(No.20070922);高等学校科技创新工程重大项目培育资金(No.70S018)资助

摘　　要：主要将aFGF基因按照植物偏爱的密码子进行基因优化,同时加上信号肽、6-组氨酸标签及凝血酶切割因子,合成全基因,再将基因插入到改造的TΩ4AB质粒中,将aFGF基因和质粒上的增强子、启动子、Ω序列及ployA加尾序列双酶切,命名为TΩ4AB-aF,将TΩ4AB-aF插入到pBI121载体中命名为pBI121-TΩ4AB-aF。利用三亲融合法将pBI121-TΩ4AB-aF转入到农杆菌菌株LBA4404中,转化苜蓿。转基因苗用卡那霉素进行筛选;并对抗性苗进行PCR、RT-PCR、Western Blot检测。结果表明,aFGF在苜蓿中得到表达。为苜蓿作为植物生物反应器奠定理论基础。After optimization of the aFGF gene for the chosen plant,the codon,the signal peptide,six-histone tag and zymoplasm cutting factor were added to the modified TΩAB plasmid,which was named TΩAB-aF after the recombination of the aFGF gene and enhancer sequence,promoter,Ω-sequence and ployA tailing sequence double digests.The binary expression vector pBI121-TΩAB-aF was constructed using TΩAB-aF and pBI121.By triparental mating the vector pBI121-TΩAB-aF was transformed into LBA4404.Medicago sativa was transformed using Agrobacterium tumefaciens,followed by selective screening and PCR,RT-PCR and Western Blot analysis.The aFGF was expressed in M.sativa.

关 键 词：AFGF 植物生物反应器 紫花苜蓿 

分 类 号：S541.9[农业科学—作物学]