蔓地亚红豆杉3′-N-去苯甲酰-2-脱氧紫杉醇苯甲酰转移酶基因的克隆与序列分析  

Cloning and Sequence Analysis of 3'-N-debenzoyltaxol N-benzoyltransferase Gene from Taxus x media

作　　者：黄仕杰[1,2] 程抒劼[1,2] 郭心悦[3] 郭丽琼[1,2] 林俊芳[1,2] 

机构地区：[1]华南农业大学食品学院,广州510640 [2]华南农业大学生物质能研究所,广州510640 [3]福建农林大学食品学院,福州350002

出　　处：《生物技术通报》2010年第12期191-195,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30671457;30871768);国家"863"资助项目(2006AA10Z301)

摘　　要：3′-N-去苯甲酰-2-脱氧紫杉醇苯甲酰转移酶(DBTNBT)是催化紫杉醇生物合成最后一步反应所需要的酶,负责将带有不完全侧链的紫杉醇前体催化形成紫杉醇。利用蔓地亚红豆杉的总DNA和总RNA为模板,采用PCR和RT-PCR技术克隆出DBTNBT基因的DNA序列和cDNA序列,测序结果显示长度分别为1465bp和1362bp,编码438个氨基酸的多肽。同源性比较分析结果表明,其碱基序列与已经报道的蔓地亚红豆杉的DBTNBT基因的一致性为99%,其氨基酸序列与已经报道的蔓地亚红豆杉的DBTNBT氨基酸序列的一致性为96%。DNA序列和cDNA序列比对发现该基因含有1个内含子。利用SWISS-PROT、DNAMAN等生物信息学工具对其蛋白序列进行了分析,为利用基因工程的方法生产紫杉醇或其前体物质提供了分子基础。3＇-N-debenzoyltaxol N-benzoyltransferase（DBTNBT）is a kind of enzyme that can catalyze the final acylation step in Taxol biosynthesis;it catalyzes the 3＇-N-debenzoyltaxol to form mature Taxol.In this study,DNA and RNA from Taxus x media were used as the template to clone the DNA sequence and cDNA sequence of DBTNBT gene.The result of sequencing showed that the length of the DNA sequence and cDNA sequence is 1 465 bp and 1 362 bp,respectively.The gene contained an opening reading frame of 1 317 bp,which coded for 438 amino acid residues.The result of alignment showed that its cDNA sequence has 99% similarity to the DBTNBT gene of Taxus x media which has been reported in NCBI,and its protein sequence has 96% similarity to DBTNBT of Taxus x media.There is one intron in this gene.Bioinformatics softwares were used to analysis the protein sequence.This research would provide the molecular base for producing Taxol and its premature materials by genetic engineering.

关 键 词：蔓地亚红豆杉 紫杉醇 3′-N-去苯甲酰-2-脱氧紫杉醇苯甲酰转移酶 基因克隆 序列分析 

分 类 号：Q943.2[生物学—植物学]