安徽农业科学2024,Vol.52Issue(21) :71-77.DOI:10.3969/j.issn.0517-6611.2024.21.015

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

陈勋 王珊 龙雨青 曾娟 周日宝 刘湘丹
安徽农业科学2024,Vol.52Issue(21) :71-77.DOI:10.3969/j.issn.0517-6611.2024.21.015

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

陈勋 1王珊 2龙雨青 3曾娟 3周日宝 3刘湘丹3
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作者信息

  • 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
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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1 245 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.

Abstract

[Objective]To clone the full-length sequence of the squalene synthase gene and perform bioinformatics and expression pattern anal-ysis 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 soft-ware,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 1 245 bp,encoding 414 amino acids.It belongs to a hydrophilic protein and is located in the cy-toplasm.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 bio-synthesis 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

Key words

Lonicera macranthoides Hand.-Mazz/Squalene synthase(SS)/Gene cloning/Bioinformatics/Expression analysis

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出版年

2024
安徽农业科学
安徽省农业科学院

安徽农业科学

影响因子:0.413
ISSN:0517-6611
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