拟南芥HSP70基因启动子表达载体的构建及在烟草BY2细胞中的应用

探讨了拟南芥的HSP70基因在液体悬浮培养的烟草BY2细胞中的表达及应用.用PCR扩增的方法从拟南芥col生态型基因组中扩增获得HSP70基因启动子序列,将其连入p1300表达载体且以GFP为报告基因,将该表达载体采用农杆菌转基因转入液体悬浮培养的烟草BY2细胞中,观察转基因细胞中报告基因GFP的表达情况.结果显示:HSP70:GFP转基因液体悬浮培养的烟草BY2细胞中有GFP的表达.该表达载体可在液体悬浮培养的BY2细胞中正常表达,且可在较短时间内获得大量实验材料,对拟南芥的HSP70基因启动子的进一步研究提供理论依据和丰富的实验材料,且可明显缩短实验周期.

Construction of the expression vector HSP70 gene promoter in Arabidopsis thaliana and application in tobacco BY2 cell

To investigate the expression and application of the HSPTO gene of arabidopsis in liquid suspension culture of BY2 cell in tobacco, using the method of PCR to amplificate genome from arabidopsis thaliana col ecotype, HSP70 gene promoter sequence, to be connected to p1300 and expression vector with GFP as report gene. The expression vector is using agrobacterium-mediated transgenic tobacco BY2 into liquid suspension culture cells to observe the expression of GFP report gene in transgenic ceils. Results ： HSWIO ： GFP transgenic tobacco BY2 of liquid suspension culture cells contains GFP. Conclusion ： the expression vector can be normal expressed in liquid suspension culture of BY2 cell. And a large number of experimental materials can be obtained in a relatively short time. HSPTO gene promoter of arabidopsis thaliana provide the oretical basis for further research and abundant experimental materials and obviously shorten the experimental period.