崔光红 王学勇 冯华 赵静雪 黄璐琦. 丹参乙酰CoA酰基转移酶基因全长克隆和SNP分析J. 药学学报, 2010,45(6): 785-790.
引用本文: 崔光红 王学勇 冯华 赵静雪 黄璐琦. 丹参乙酰CoA酰基转移酶基因全长克隆和SNP分析J. 药学学报, 2010,45(6): 785-790.
CUI Guang-Gong, WANG Hua-Yong, FENG Hua, DIAO Jing-Xue, HUANG Lu-Qi. Molecular cloning and SNP analysis of a acetyl-CoA C-acetyltransferase gene (SmAACT) from Salvia miltiorrhizaJ. 药学学报, 2010,45(6): 785-790.
Citation: CUI Guang-Gong, WANG Hua-Yong, FENG Hua, DIAO Jing-Xue, HUANG Lu-Qi. Molecular cloning and SNP analysis of a acetyl-CoA C-acetyltransferase gene (SmAACT) from Salvia miltiorrhizaJ. 药学学报, 2010,45(6): 785-790.

丹参乙酰CoA酰基转移酶基因全长克隆和SNP分析

Molecular cloning and SNP analysis of a acetyl-CoA C-acetyltransferase gene (SmAACT) from Salvia miltiorrhiza

  • 摘要:

    乙酰CoA酰基转移酶 (acetyl-CoA C-acetyltransferase, AACT) 为萜类化合物生物合成甲羟戊酸 (MVA) 途径的起始酶, 催化使2个分子的乙酰CoA缩合为乙酰乙酰CoA。利用基因芯片结合RACE方法, 从丹参毛状根中得到一个AACT基因 (SmAACT, accession No. EF635969), 该基因全长1 623 bp, 含有11 200 bp的开放阅读框 (open reading frame, ORF), 编码399个氨基酸, 对应基因组序列含有9个内含子。序列同源性分析和分子进化分析表明SmAACT是一种与植物类异戊二烯MVA途径相关的蛋白, 其表达量受到生物和非生物诱导子酵母和Ag+的诱导, 并伴随丹参酮类成分积累。在丹参根、茎、叶中均有表达, 根中的表达量明显高于茎和叶中的表达量。单核苷酸多态性分析表明, 在第6至第9内含子约600 bp范围内, SmAACT共存在33个多态位点, 且表现出产地特异基因型。该基因的克隆分析为研究丹参酮类成分的次生代谢调控机制奠定了基础。

     

    Abstract:

    Acetyl-CoA C-acetyltransferase (AACT) is the first enzyme in the terpene synthesis pathway, catalyzed two units of acetyl-CoA to acetoacetyl-CoA.  In order to study the tanshinone biosynthesis in Salvia miltiorrhiza, a novel AACT gene, SmAACT, was cloned using cDNA microarray and RACE strategy.  The full length cDNA of SmAACT is 1 623 bp (accession No. EF635969), which contained a 1 200 bp open reading frame (ORF) encoding a 399 amino acid protein.  Nine introns were found in the genomic sequence.  SmAACT was upregulated by YE and Ag+ elicitors both with cDNA microarray and quantitative RT-PCR analyses along with the accumulation of tanshinones.  Sequence homology comparison and phylogenetic analysis all suggested that SmAACT belonged to the class of acetyl-CoA C-acetyltransferase.  The transcription level of SmAACT was  relatively higher in root than that in stem and leaf tissues.  SNP analysis revealed that SmAACT was highly variable in the region of 6 to 9 introns with 33 SNPs in the 600 bp region, there are 5 SNPs in the cDNA region while they are all synonymous cSNPs.  Some special genotypes were found in Salvia miltiorrhiza from different areas.  SmAACT will be an useful gene for further analyze the mechanism of gene regulation among the tanshinones biosynthesis.

     

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