Abstract:

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.

Key words:  Artemisia annua L., artemisininin, cyp71av1 gene, cpr gene

摘要：

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

GenBank中青蒿紫穗槐二烯氧化酶（CYP71AV1）和细胞色素P450还原酶（CPR）的基因序列设计特异引物,从青蒿cDNA中扩增出了cyp71av1和cpr

基因片段,分别连上CaMV 35S启动子和NOS终止子,然后将cyp71av1和cpr基因的表达盒依次插入到植物表达载体pCAMBIA2300中,获得含有

cyp71av1和cpr基因的植物表达载体p2300-GYR。通过农杆菌介导法转化青蒿,获得了20株转基因植株,PCR检测以及Southern杂交分析证实外源基

因已被成功地导入青蒿基因组中。采用HPLC-ELSD测定转基因青蒿中青蒿素的含量,结果表明,大部分转基因青蒿中青蒿素的含量高于对照植株,

其中,转基因青蒿中青蒿素含量最高约为对照的2.4倍。该研究证明过量表达cyp71av1和cpr基因是提高青蒿中青蒿素含量的有效方法。

关键词: 青蒿；青蒿素； cyp71av1基因； cpr基因