黄连和三角叶黄连去甲乌药碱合成酶(NCS)基因克隆与原核表达分析  

Cloning and prokaryotic Expression analysis of noraconine synthetase(NCS)gene in Coptis chinensis Franch.and Coptis deltoidea C.Y.Cheng et Hsia

作　　者：李懿柔 柯汶佳 郑琪 李子涵 周涛 黄玲 马云桐[1] LI Yirou;KE Wenjia;ZHENG Qi;LI Zihan;ZHOU Tao;HUANG Ling;MA Yuntong(School of Pharmacy,State Key Laboratory of Southwest Chinese Medicine Resources,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,Sichuan)

机构地区：[1]成都中医药大学药学院、西南特色中药资源国家重点实验室,四川成都611137

出　　处：《中药与临床》2024年第4期5-10,31,共7页Pharmacy and Clinics of Chinese Materia Medica

摘　　要：目的:探究黄连和三角叶黄连去甲乌药碱合成酶(NCS)基因的表达差异,为理解同属植物生物碱含量差异机制奠定基础。方法:采集相同生长年限的黄连和三角叶黄连,分析其叶片、根和根茎的生物碱含量和生物碱合成的关键基因表达量,对关键基因进行克隆并构建原核表达载体。结果:黄连和三角叶黄连各部位生物碱含量存在差异,且NCS基因与各生物碱含量关系最密切,生物信息学分析显示NCS基因在两种黄连中无明显差异,同时亲缘关系十分接近;成功构建NCS基因的原核表达载体。结论:黄连和三角叶黄连的NCS基因是生物碱合成的关键基因,其在根的表达差异可能是影响生物碱合成的重要因素。Objective:To explore the difference in the expression of noraconitine synthetase(NCS)gene between Coptis chinensis Franch.(Huanglian)and Coptis deltoidea C.Y.Cheng et Hsiao(Sanjiaoye Huanglian),and to lay a foundation for understanding the mechanism of alkaloid content difference in plants of the same genus.Methods:Huanglian and Sanjiaoye Huanglian with the same growth years were collected.The contents of alkaloids and the expression of key genes involved in alkaloid synthesis in leaves,roots and rhizome in these two plants were analyzed.The key genes were cloned and the prokaryotic expression vector was constructed.Results:There were differences in the alkaloids content in various parts of Huanglian and Sanjiaoye Huanglian,and the NCS gene was most closely related to the alkaloids content.The bioinformatics analysis showed that there was no significant difference in the NCS gene between the two plants,and the homology was very close.The prokaryotic expression vector of NCS gene was successfully constructed.Conclusion:The NCS gene of Huanglian and Sanjiaoye Huanglian is the key gene in alkaloid synthesis,and the difference of its expression in roots may be an important factor affecting alkaloid synthesis.

关 键 词：黄连 三角叶黄连 去甲乌药碱合成酶 基因克隆 表达分析 

分 类 号：R282[医药卫生—中药学]