荔枝类黄酮糖基转移酶(UFGT)基因的克隆及其原核表达研究  

Cloning Glucose-flavonoid 3-o-glucosyltransferase(UFGT) from Litchi and Expression in Escherichia coli

作　　者：赵志常[1,2] 胡福初[1,2] 胡桂兵[1,2] 王惠聪[1,3] 杨转英[1,2] 苏纯兰[1,2] 李加强[1,2] 

机构地区：[1]华南农业大学园艺学院果树种质资源与品种改良研究室,广东广州510642 [2]华南农业大学园艺生物技术研究所,广东广州510642 [3]华南农业大学园艺学院南方果树生理研究室,广东广州510642

出　　处：《广西师范大学学报（自然科学版）》2011年第4期104-110,共7页Journal of Guangxi Normal University:Natural Science Edition

基　　金：国家科技支撑计划(2006BAD01A1705);现代农业产业技术体系建设专项资金资助项目(nycytx-32);国家自然科学基金资助项目(30971985)

摘　　要：类黄酮糖基转移酶(UFGT)可以把不稳定的花色素催化成花色素苷,是花色素苷合成过程中的最后一个酶。在一定范围内,类黄酮糖基转移酶活性与花色素苷的合成呈现正相关,抑制UFGT酶活性比抑制其他酶更能影响花色素苷的合成。很多研究结果表明,UFGT是花色素苷合成的关键酶基因。本研究根据在荔枝上已知的UFGT基因片段,采用3′RACE和5′RACE技术克隆得到UFGT的全长cDNA。采用特异引物扩增得到该基因的基因组DNA的全长序列,发现该基因含有一个内含子。将该基因全长cDNA序列构建到原核表达载体pET32a中,通过1 mmol/L IPTG诱导表达,得到大约60 KDa的融合蛋白。Litchi is native to South China＇s tropical fruit trees,the fruit color is various types. Function of Glucose-flavonoid 3-o-glucosyltransferase （UFGT） is catalyzed the unstable anthocyanidin into antho-cyanin. It is the last enzyme in anthocyanin biosynthetic pathway. Within a certain range,UFGT activity is positively related to anthocyanin synthesis,and enzyme activity of UFGT inhibited has more influence on anthocyanin synthesis than other enzymes. Many research results show that UFGT is a key enzyme gene in anthocyanin synthesis. This study is based on the known UFGT gene fragments in the Litchi,the full-length cDNA of UFGT was cloned using the 3tRACE and 5＇RACE methods. It was found an intron in the genomic DNA sequence compared to cDNA sequence. The cDNA fragment of UFGT directionally cloned into the vector pET32a,the UFGT fusion protein about 60 KDa was expressed by induction I mmol/L IPTG in E. coll.

关 键 词：荔枝 花色素苷 类黄酮糖基转移酶 基因克隆 原核表达 

分 类 号：S667.1[农业科学—果树学]