北沙参补骨脂素合成酶基因 GlPS1 原核表达载体的构建和蛋白诱导表达  

Construction of Prokaryotic Expression Vector for Psoralen SynthaseGene GlPS1 from Glehniae Radix and Its Protein Induced Expression

作　　者：张馨方 宋洁洁 徐瑶 田雪梅[1] 谭玲玲[1] 高婷[1] ZHANG Xinfang;SONG Jiejie;XU Yao;TIAN Xuemei;TAN Lingling;GAO Ting(Key Laboratory of Plant Biotechnology in Universities of Shandong Province/College of Life Science,Qingdao Agricultural University,Qingdao 266109,China;The Second Middle School of Juancheng County,Heze 274600,China)

机构地区：[1]青岛农业大学生命科学学院/山东省高校植物生物技术重点实验室,山东青岛266109 [2]山东省鄄城县第二中学,山东菏泽274600

出　　处：《青岛农业大学学报（自然科学版）》2023年第2期118-123,共6页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家自然科学基金(81903748)。

摘　　要：北沙参(Glehniae Radix)为伞形科植物珊瑚菜(Glehnia littoralis)的干燥根,具有良好的药用价值和较高的经济价值。补骨脂素(psoralen)是北沙参香豆素类化合物主要的活性成分之一,对白血病细胞有较强的杀伤作用,对支气管平滑肌有舒张作用。补骨脂素合成酶(GlPS)是北沙参补骨脂素生物合成途径的最后一步关键酶。本试验通过PCR扩增获得对北沙参补骨脂素合成酶基因GlPS1,通过酶切、连接、转化、平板培养和提取质粒构建原核表达载体;将构建好的表达载体转入大肠杆菌BL21(DE3)中,实现GlPS1基因的IPTG诱导原核表达。该研究表明被诱导表达的GlPS1蛋白存在于菌体沉淀中,不溶于上清液。研究结果为验证GlPS1基因的功能及使用基因工程方法调控GlPS1蛋白表达奠了基础。Glehniae Radix,the dried root of an Umbellales plants Glehnia littoralis,has highly medicinal and economic value.Psoralen,as one of the main active components of coumarins in Glehniae Radix,exerts important function in killing leukemia cell and inducing relaxation in bronchial smooth muscle.Psoralen synthase(GlPS)is a key enzyme that catalyzes the last step of the psoralen biosynthesis in Glehniae Radix.In this study,psoralin synthase gene GlPS1 was obtained by PCR amplification,and the prokaryotic expression vector was constructed by enzyme digestion,ligation,transformation,plate culture and plasmid extraction.Then the constructed vector was transfected into Escherichia coil BL21(DE3),by which GlPS1 protein was expressed in prokaryotic cells with IPTG induction.It was found that the expressed GlPS1 protein existed in bacterial precipitation and remained undissolved in bacterial precipitation.The findings lay the foundation for verifying the function of the GlPS1 gene and regulating the expression of GlPS1 protein using genetic engineering methods.

关 键 词：北沙参 GlPS1基因 原核表达载体构建 蛋白诱导表达 

分 类 号：Q943.2[生物学—植物学]