雷公藤单萜合酶基因TwMS的克隆及蛋白表达分析

发布时间：2018-03-20 02:03

本文选题：雷公藤　切入点：单萜合酶　出处：《中国中药杂志》2017年07期 　论文类型：期刊论文

【摘要】：该研究克隆得到1条雷公藤单萜合酶基因TwMS。其完整开放阅读框为1797bp,编码579个氨基酸,相对分子质量为69.75kDa、理论等电点为5.37。生物信息学分析表明,TwMS具有单萜合酶的特征结构域,属于萜类合酶TPSb亚家族。实时荧光定量PCR检测显示,经茉莉酸甲酯(MeJA)诱导后,TwMS的相对表达量显著上调,并在24 h达到最高。此外,该研究在大肠杆菌BL21(DE3)中成功表达TwMS蛋白,为进一步研究该基因的功能奠定了基础。
[Abstract]:In this study, a monoterpene synthase gene TwMSfrom Tripterygium wilfordii was cloned. Its complete open reading frame was 1797bp, encoding 579 amino acids, the relative molecular weight was 69.75kDa, and the theoretical isoelectric point was 5.37.The bioinformatics analysis showed that TwMS had the characteristic domain of monoterpene synthase. It belongs to the TPSb subfamily of terpene synthase. Real-time quantitative PCR analysis showed that the relative expression of TwMS was significantly up-regulated after induction by methyl jasmonate, and reached the highest level in 24 h. In addition, the TwMS protein was successfully expressed in E. coli BL21DE3. It lays a foundation for further study of the function of the gene.
【作者单位】：首都医科大学中医药学院中医络病研究北京重点实验室;中国中医科学院中药资源中心道地药材国家重点实验室培育基地;
【基金】：国家自然科学基金优秀青年科学基金项目(81422053);国家自然科学基金面上项目(81373906)
【分类号】：S567.19

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