基于基因枪技术的拟南芥瞬时表达转化方法的建立

Setup of Transient Expression Assay System Based on Particle Bombardment in Arabidopsis

以拟南芥叶片为材料,分析不同轰击参数对基因枪转化后的叶片中LUC表达效率的影响.结果表明,-0.09 MPa的真空度4、.5 MPa可裂膜压力以及5 cm的载体膜与叶片距离相结合能够获得最佳的转化效率.同时,研究ABA对基因枪转化后的叶片中LUC基因表达的影响,结果显示转化后的叶片具有正常的生理活性,ABA处理能够较好地诱导LUC基因的表达.

In this research, the authors detected the luciferase activity in particle bombardment transformed Arabidopsis leaves with different bombard parameters, and found that with -0.09 MPa vacuum, 4.5 MPa pressure and 5 cm target distance could obtain highest transform rate. The authors also compared the LUC activity in control and ABA treatment drived by four repeated ABRE elements. The results indicated that ABA treatment enhanced LUC activity obviously, these results suggested that transformed leaves maintain normal biological activity and are suitable for transient expression assay.