过表达长春花JAR1基因促进文朵灵和长春质碱的生物合成

发布时间：2018-05-08 01:01

本文选题：长春花 + 长春质碱和文朵灵　；参考：《生物技术通报》2017年06期

【摘要】：通过克隆长春花JAR1基因并在长春花叶片中瞬时过表达,研究该基因对文朵灵及长春质碱生物合成的调控。以总c DNA为模板,克隆JAR1基因,并构建重组质粒p BIGD-JAR1。经农杆菌介导瞬时转化长春花叶片后,利用半定量RT-PCR与高效液相分析JAR1过表达对文朵灵及长春质碱合成的影响。结果显示,成功从长春花叶片中克隆1 700 bp左右的JAR1基因;瞬时转化p BIGD-JAR1的长春花叶片中JAR1基因的转录水平高于空载体对照;JAR1的过表达诱导了合成途径中关键酶基因Cr TDC、Cr G10H、Cr DL7H、Cr STR、Cr LAMT的表达,以及文朵灵和长春质碱的积累。结合实验结果分析,JAR1基因过表达可能通过促进茉莉酸(JA)信号转导途径来诱导关键代谢途径酶基因的表达进而促进长春花中文朵灵和长春质碱的积累。
[Abstract]:The regulation of JAR1 gene on the biosynthesis of Vindolin and vincristine was studied by cloning and transient overexpression in Changchun flower leaf tablets. Using total c DNA as template, the JAR1 gene was cloned and the recombinant plasmid pBIGD-JAR1 was constructed. After transient transformation of rosebush leaves mediated by Agrobacterium tumefaciens, the effects of JAR1 overexpression on the synthesis of vincristine and vincristine were analyzed by semi-quantitative RT-PCR and high performance liquid chromatography (HPLC). The results showed that 1 700 BP JAR1 gene was successfully cloned from the leaves of Catharanthus roseus. The transcriptional level of JAR1 gene in the leaves of Catharanthus roseus instant transformed with p BIGD-JAR1 was higher than that of the blank vector control JAR1. The overexpression of JAR1 gene induced the expression of the key enzyme gene Cr TDCN, Cr G10H, Cr DL7H, and the accumulation of vendolin and vincristine in the biosynthesis pathway. Combined with the experimental results, the overexpression of JAR1 gene may induce the expression of key metabolic pathway enzyme genes by promoting JAA signal transduction pathway, thus promoting the accumulation of VCR and vincristine.
【作者单位】：大连工业大学生物工程学院;
【基金】：辽宁省教育厅基本科研业务项目(2016J048)
【分类号】：Q943.2;S567.239

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