灵芝倍半萜合酶基因的克隆及表达  

Cloning and expression of the sesquiterpene synthase gene from Ganoderma lucidum

作　　者：王丽芝[1] 浦香东 谭世强[2] 孙思杰[1] 毕钰彤 孙超[3] 陈士林[3] 王海英[1] WANG Li-zhi PU Xiang-dong TAN Shi-qiang SUN Si-jie BI Yu-tong SUN Ohao OHEN Shi-lin WANG Hai-ying(School of Traditional Chinese Pharmacy, Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China Department of Experimental Teaching,Tianjin University of Traditional Chinese Medicine, Tianjin 300193, China Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences （CAMS） and Peking Union Medical College, Beijing 100193, China)

机构地区：[1]天津中医药大学中药学院,天津300193 [2]天津中医药大学实验教学部,天津300193 [3]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《河北农业大学学报》2017年第2期67-72,共6页Journal of Hebei Agricultural University

基　　金：国家自然科学基金(81603221);天津市自然科学基金(16JCQNJC09700);天津市高等学校科技发展基金(20120202/20140209);大学生创新创业训练计划项目(201610063003)

摘　　要：倍半萜合酶是倍半萜化合物生物合成途径中的关键酶。本试验从灵芝基因组数据库中筛选出1个编码灵芝萜类合酶的基因,并对其进行克隆及表达。序列分析表明,该基因开放阅读框的长度为1041bp,编码346个氨基酸;成功构建了1个灵芝萜类合酶基因的原核表达载体,并完成了该基因编码的倍半萜合酶在大肠杆菌中的可溶性表达;顶空固相微萃取—气相色谱质谱法分析结果表明重组的灵芝萜类合酶具有催化活性。为后续研究萜类化合物的多样性奠定基础。Sesquiterpene synthases are key enzymes involved in the biosynthetic pathway of sesquiterpenoids. In this study, the candidate gene encoding a sesquiterpene synthase was ob- tained from Ganoderrna lucidum genome database, and then was cloned and expressed in E. coll. The result of sequence analysis showed that the gene contained a complete open reading frame（ORF）with 1041 bp, encoding 346 amino acids. A prokaryotic expression vector of the sesquiterpene synthase was successfully constructed, and the recombinant sesquiterpene syn- thase was successfully expressed in E. coli BL 21. The result of solid-phase micro-extraction and GC-MS （SPME-GC-MS） revealed that the recombinant sesquiterpene synthase had the catalytic activity. These results laid foundation for the follow-up investigation of sesquiterpe- noids diversity.

关 键 词：灵芝 倍半萜合酶 生物合成 

分 类 号：S567.31[农业科学—中草药栽培]