灰毡毛忍冬SS基因的克隆及表达分析  

Cloning and Expression Analysis of Squalene Synthase Gene in Lonicera macranthoides

作　　者：陈勋 王珊[2] 龙雨青 曾娟 周日宝[3] 刘湘丹[3] CHEN Xun;WANG Shan;LONG Yu-qing(Affiliated Nanhua Hospital,University of South China,Hengyang,Hunan 421002;The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,Hunan 410005;School of Pharmacy,Hunan University of Chinese Medicine,Changsha,Hunan 410208)

机构地区：[1]南华大学附属南华医院,湖南衡阳421002 [2]湖南中医药大学第二附属医院,湖南长沙410005 [3]湖南中医药大学药学院,湖南长沙410208

出　　处：《安徽农业科学》2024年第21期71-77,共7页Journal of Anhui Agricultural Sciences

基　　金：湖南省自然科学基金项目(2021JJ30497);湖南省教育厅资助科研项目(16B193);湖南省自然科学基金联合项目(2024JJ8221);湖南省中医药科研计划项目(D2022134);湖南省教育厅科学研究项目(21C0245)。

摘　　要：[目的]克隆灰毡毛忍冬SS基因全长序列,并进行生物信息学和表达模式分析。[方法]提取灰毡毛忍冬花总RNA,通过RT-PCR和RACE技术克隆LmSS基因的全长cDNA序列;运用相关软件对该基因序列进行生物信息学分析;利用qRT-PCR测定灰毡毛忍冬茎、叶和不同花期花中该基因的相对表达量。[结果]克隆的LmSS基因开放阅读框(ORF)长度为1245 bp,编码414个氨基酸,属于亲水性蛋白,定位于细胞质中,具有典型的多聚异戊二烯基合成酶活性结构域,与其他植物SS基因具有较高同源性。LmSS基因在灰毡毛忍冬不同花期和器官中表达有组织特异性。[结论]该研究成功地克隆灰毡毛忍冬中的SS基因,为进一步研究该基因的功能奠定基础,同时为探究灰毡毛忍冬皂苷生物合成和调节机制提供了科学依据。[Objective]To clone the full-length sequence of the squalene synthase gene and perform bioinformatics and expression pattern analysis from Lonicera macranthoides.[Method]Total RNA was extracted from Lonicera macranthoides,and the full-length cDNA sequence of LmSS gene was cloned using RT-PCR and RACE techniques,bioinformatics analysis was conducted on the gene sequence using relevant software,the relative expression of the gene in stem,leaf,and different flower period was determined by using Real-time PCR.[Result]The open reading frame(ORF)of LmSS gene was 1245 bp,encoding 414 amino acids.It belongs to a hydrophilic protein and is located in the cytoplasm.LmSS gene has a typical polyisoprene synthase active domain,which has high homology with other plant SS genes.The expression of LmSS gene is tissue-specific in different flowering stages and organs of Lonicera macranthoides.[Conclusion]This study successfully cloned the SS gene in Lonicera macranthoides,laying a foundation for further research on its function and providing a research basis for exploring the biosynthesis and regulatory mechanism of saponins in Lonicera macranthoides.

关 键 词：灰毡毛忍冬 鲨烯合酶(SS) 基因克隆 生物信息学 表达分析 

分 类 号：S567.7[农业科学—中草药栽培]