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.
出处
《高技术通讯》
CAS
CSCD
北大核心
2007年第4期424-429,共6页
Chinese High Technology Letters
基金
863计划（2002AA212071）资助项目.
