太子参鲨烯环氧酶基因的克隆及其表达分析

发布时间：2018-03-03 08:45

本文选题：太子参　切入点：三萜皂苷　出处：《中草药》2017年13期 　论文类型：期刊论文

【摘要】：目的对太子参三萜类皂苷生物合成关键调控酶鲨烯环氧酶1(SQE1)基因进行全长c DNA克隆和功能分析。方法基于其他植物SQE基因的同源序列设计简并引物,以太子参块根总RNA为模板,RT-PCR结合RACE技术克隆太子参SQE1基因的全长c DNA,并进行生物信息学分析。利用农杆菌叶盘转化法将SQE1基因转化烟草,研究其对烟草总三萜量的影响。结果获得太子参SQE1基因全长c DNA序列2 038 bp,该序列包含1 554 bp的开放阅读框,编码517个氨基酸,相对分子质量为5.67×104,等电点为8.8,与其他药用植物的SQE蛋白具有较高的同源性,含有FAD结合结构域和4个跨膜区域,转太子参SQE1基因的烟草的总三萜量明显高于非转基因植株。结论首次克隆获得太子参SQE1基因的全长c DNA,该基因的异源表达可一定程度提高转基因植物的总三萜量,为阐明与应用太子参三萜类成分生物合成途径提供科学依据。
[Abstract]:Objective to clone and analyze the full-length c DNA of squalene cyclooxygenase 1 (squalene cyclooxygenase 1) gene, a key regulator of triterpenoid saponins biosynthesis of Pseudostellaria heterophylla. Methods degenerate primers were designed based on the homologous sequence of SQE gene in other plants. The full-length cDNA of SQE1 gene of Pseudostellariae heterophylla was cloned by RT-PCR and RACE using total root RNA of P. princeliae as template. Bioinformatics analysis was carried out. The SQE1 gene was transformed into tobacco by Agrobacterium tumefaciens transformation method. Results the full-length c DNA sequence of Radix Pseudostellariae SQE1 gene was 2 038 BP, which contained an open reading frame of 1 554 BP, encoding 517 amino acids. The relative molecular weight is 5.67 脳 10 ~ 4 and the isoelectric point is 8.8. It has high homology with other medicinal plants' SQE protein, and contains FAD binding domain and four transmembrane domains. Conclusion the total triterpenoids of the transgenic plants were significantly higher than that of the non-transgenic plants. Conclusion the full-length c-DNA of the SQE1 gene was obtained for the first time, and the heterologous expression of the gene could improve the total triterpenoid quantity of the transgenic plants to some extent. It provides scientific basis for elucidation and application of biosynthesis pathway of triterpenoids of Pseudostellaria heterophylla.
【作者单位】：福建农林大学生命科学学院;
【基金】：国家自然科学基金资助项目(30900915) 福建省教育厅资助高校重点项目(JK2012012)
【分类号】：S567.53

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