雷公藤鲨烯合酶基因全长cDNA克隆及诱导表达分析
Cloning and expression analysis of squalene synthase gene in Tripterygium wilfordii
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摘要: 根据雷公藤悬浮细胞转录组数据设计引物,克隆得到雷公藤鲨烯合酶基因TwSQS全长cDNA (GenBank:KR401220),并对其进行生物信息学分析和诱导表达分析。TwSQS cDNA全长为1800 bp,开放阅读框为1242 bp,编码413个氨基酸,编码的蛋白理论等电点为7.94,相对分子质量为47.20 kD。雷公藤悬浮细胞经茉莉酸甲酯(MJ)诱导后,荧光定量PCR表明TwSQS相对表达量明显上升,并于诱导后4 h达到最高。本研究首次克隆得到雷公藤SQS基因,为进一步解析雷公藤三萜类成分生物合成途径奠定基础。Abstract: In this paper, we cloned the full-length cDNA of TwSQS from Tripterygium wilfordii suspension cells (GenBank:KR401220) and performed the bioinformation and mRNA expression analysis. The expression after methyl jasmonate (MJ) treatment of the gene was detected by RT-PCR. The full-length cDNA of TwSQS was 1800 bp containing a 1242 bp open reading frame (ORF) encoding a polypeptide of 413 amino acids. The theoretical isoelectric point (pI) was 7.94 and the calculate molecular weight was about 47.20 kD. The relative expression level of TwSQS was deduced by MJ and reached the highest at 4 h after the treatment. The gene information we got in this study enriched the biosynthesis pathway of triterpenoids in Tripterygium wilfordii and laid foundation for further studies.
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