莴苣1-FFT基因的克隆及其功能分析  

Cloning and functional analysis of fructan： fructan 1-fructosyltransferase from Lactuca sativa

作　　者：李慧娟[1] 张学成[1] 岳桂东[2] 张举仁[2] 

机构地区：[1]中国海洋大学生命科学与技术学部,青岛266003 [2]山东大学生命科学学院,济南250100

出　　处：《高技术通讯》2007年第4期424-429,共6页Chinese High Technology Letters

基　　金：863计划（2002AA212071）资助项目.

摘　　要：利用反转录PCR技术和cDNA末端快速扩增（RACE）技术从莴苣中分离出1-FFT（果聚糖：果聚糖1-果糖基转移酶）候选全长cDNA,序列分析表明该基因的开放阅读框中具有β-果糖苷酶单元（蔗糖结合框）、RDP域和EC域三个活性中心域。Southern杂交分析表明该基因在莴苣基因组中以单拷贝形式存在。利用农杆菌介导的叶盘转化法将该基因转入无1-FFT基因的烟草细胞,获得了转基因植株。体外酶反应证实转基因叶片提取物中具有1-FFT活性,该基因编码1-FFT。The full-length cDNA encoding fructan： fructan 1-fructosyltransferase 1-FFT from Lactuca sativa was cloned by RT- PCR and RACE method. Its deduced protein shares three highly conserved regions including β-fructosidase region （sucrose-binding box）, RDP region and EC region with other fructosyltransferases. Southern blot analysis revealed that 1-FFF was encoded by single-copy gene. Transgenic tobacco was obtained by Agrobacterium-mediated tobacco leaf disc transformation protocol. The results of fructan synthesis in vitro suggested that the extracts of transgenic tobacco leaves displayed fructan： fructan 1-fructosyltransferse activity. It could be inferred that the full-length 1-FFT cDNA encoded a functional fructan： fructan 1-fructosyltransferse.

关 键 词：莴苣 RACE 果聚糖:果聚糖1-果糖基转移酶基因 转基因烟草 

分 类 号：S636.2[农业科学—蔬菜学]