过量表达cyp71av1和cpr基因提高青蒿中青蒿素的含量

青蒿素是从青蒿植株地上部分提取的一种含有过氧桥结构的倍半萜内酯,是目前最有效的治疗疟疾的药物。为了提高青蒿中青蒿素的含量,根据

Over-expressing cyp71av1 and cpr Genes Enhances Artemisinin Content in Artemisia annua L.

Artemisinin, a sesquiterpene lactone endoperoxide derived from plant Artemisia annua L., is the most effective anti-malarial drug. In an effort to increase the artemisinin production, cyp71av1 and cpr genes were cloned from A. annua cDNA using two pairs of gene-specific primers which were designed according to the gene sequences at GenBank. Both genes were inserted between cauliflower mosaic virus (CaMV) 35S promoter and NOS terminator to construct expression cassettes, which were then cloned into plant expression vector pCAMBIA2300, resulting in p2300-GYR containing both cyp71av1 and cpr genes. Agrobacterium-mediated method was used to transform A. annua, and 20 independent transgenic plants were obtained. PCR detection and Southern blot analysis demonstrated that the foreign genes were successfully introduced into A. annua genome. Artemisinin contents in transgenic plants were determined by HPLC-ELSD and the result showed that artemisinin contents in most transgenic plants were higher than that in control plants, with the highest artemisinin content in transgenic plant being about 2.4-fold to that in the control plant. This study demonstrates that over-expressing cyp71av1 and cpr genes is an effective approach to enhance artemisinin content in Artemisia annua L.