人RANTES基因在转基因青蒿植株中表达的测定

Assay of human RANTES gene expressed in transgenic plants of Artemisia annua

目的 为了增强青蒿的特异药理活性,探讨人β-趋化因子RANTES基因在青蒿细胞中表达的可能性。方法 将克隆的RANTES基因经Ti质粒衍生的双元表达载体pROKII导入根癌农杆菌LBA4404菌株,通过叶盘共培养法已获得一批表现卡那霉素抗性的转基因青蒿植株。结果 PCR和Southern印迹杂交分析表明,RANTES基因已导入并整合到青蒿基因组中;RT-PCR、Northern印迹、Western印迹杂交结果证实,RANTES基因已在转基因青蒿植株中成功表达。结论 首次报道了RANTES基因在转基因青蒿植株中的表达,为基因工程改造青蒿打下了良好的基础。

Object To enhance the specific pharmacological activity of A. annua and explore the possibility for expression of human β-chemokine RANTES gene in A. annua cells. Methods The cloned RANTES gene was introduced into Agrobacterium tumefaciens LBA4404 strain by Ti plasmid-derived binary vector pPOKII. A batch of transgenic A. annua plants with kanamycin resistance was regenerated by disk cocultivation procedure. Results The assay by polymerase chain reaction (PCR) and Southern blotting showed that RANTES gene had introduced and integrated into the genome of A. annua. The results of RT-PCR, Northern blotting, and Western blotting confirmed that RANTES gene had expressed successfully in transgenic A. annua plants. Conclusion The expression of RANTES gene in transgenic A. annua plants, which pave a direct path to modify A. annua by genetic engineering.