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2025年4月5日 星期六
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金龙胆草查尔酮合酶基因(CHS)的克隆及其在花期不同组织的表达量分析
刘姗1,孙蓉2,唐自钟1,郑天润2,高静雷2,陈惠1,李成磊2
1. 四川农业大学
2. 四川农业大学生命科学与理学院
Cloning of Chalcone Synthase Gene (CHS) from Conyza blinii Lévl. and It's Expression Analysis in Different Tissues During Florescence
2, 2, 2, 2,hui chen3, 2
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摘要 查尔酮合酶(chalcone synthase, CHS)是植物类黄酮化合物合成的第一关键酶。为了获得金龙胆草(Conyza blinii Lévl.)CHS基因,本研究采用快速扩增cDNA末端(rapid-amplification of cDNA ends, RACE)技术克隆得到了金龙胆草CHS基因的DNA和cDNA全长序列,并进行生物信息学分析,通过RT-PCR检测该基因在花期不同组织部位中的表达情况并检测对应部位的相对花青素含量。结果表明,获得的CHS基因DNA全长2 098 bp,包括2个内含子;cDNA全长为1 692 bp,含有完整开放阅读框(1 197 bp),编码399个氨基酸,包含查尔酮合酶的标志性序列,命名为CbCHS(GenBank登录号: KJ155749)。序列比对结果显示,金龙胆草CbCHS基因与已报道的其它植物的CHS氨基酸序列具有很高相似性(最高为95%)。半定量RT-PCR结果显示,在叶和茎的表达量最高,相对表达量分别达到87.03 %和98.33 %,表达量之间没有显著差异(P>0.05);在根中CbCHS基因表达量最少,相对表达量仅为9.43 %,表达量与其余部位具有显著差异(P<0.05)。花青素含量也以茎和叶中最高,根中微量,表明CbCHS基因与花青素的蓄积有一定相关性。本研究首次克隆了金龙胆草CbCHS基因的全长DNA及cDNA序列,研究了其在花期不同组织器官中的表达量差异及与花青素的含量关系,为进一步研究CbCHS基因对金龙胆草黄酮含量的影响及其调控机制提供了基础资料。
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刘姗
孙蓉
唐自钟
郑天润
高静雷
陈惠
李成磊
关键词 金龙胆草查尔酮合酶(CHS)克隆生物信息学分析表达量分析    
Abstract:Chalcone synthase (CHS) is a pivot for the biosynthesis of flavonoid antimicrobial phytoalexins and anthocyanin pigments in plants. To clone CHS gene from Conyza blinii Lévl. and analyze its expression in different tissues during florescence, the full- length DNA cloned by RACE was 2 098 bp, including 2 introns (486 and 415 bp); the coding region of cDNA consisted of 1 692 nucleotides, and predicted to code a protein of 399 amino acids which included all the active sites of CHS, named CbCHS (GenBank accession No. KJ155749). Bioinformatic analysis result showed that CbCHS amino acid sequence had a high homology with other opened CHS genes in plants (the highest value of 95%). The RT-PCR result showed that CbCHS was highly expressed in stems (87.03%) and leaves (98.33%) , showed no significant difference between them (P>0.05), while roots had the lowest expression (9.43%), and showed a significant difference with other tissues (P<0.05). The anthocyanin content was the highest in stems and leaves, and the lowest in roots, which indicated that there was some relevance between the CbCHS expression and anthocyanins accumulation. The DNA and cDNA sequence of CbCHS gene were firstly obtained from Conyza blinii Lévl. with the research of relationship between the anthocyanin content and gene espression in different tissues of Conyza blinii Lévl.. The results of this study provide some technology basis for the key enzyme expression and regulation mechanism analysis in effective components biosynthesis pathway of Conyza blinii Lévl..
Key wordsConyza blinii Lévl.    Chalcone synthase    Clone    Bioinformatic analysis    Expression analysis
收稿日期: 2013-12-04     
通讯作者: 陈惠   
引用本文:   
刘姗1,孙蓉2,唐自钟1,郑天润2,高静雷2,陈惠1,李成磊2. 金龙胆草查尔酮合酶基因(CHS)的克隆及其在花期不同组织的表达量分析[J]. , 2014, 22(6): 703-711.
2, 2, 2, 2,hui chen3, 2. Cloning of Chalcone Synthase Gene (CHS) from Conyza blinii Lévl. and It's Expression Analysis in Different Tissues During Florescence. , 2014, 22(6): 703-711.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I6/703
 
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