用基因枪法将bar基因导入杜氏盐藻及转基因藻株的检测

目的：探讨基因枪转化方法不同轰击条件和启动子的选择对转化杜氏盐藻的影响。方法：以抗除草剂(bar)基因作为报告基因，用基因枪法将其导入杜氏盐藻。通过草丁膦筛选培养获得转化藻株，对转化藻株进行分析。结果：在氦气压力为100L／min条件下，微弹轰击2次比微弹轰击1次或3次的效果都好；杜氏盐藻碳酸酐酶(CA1)基因启动子可驱动bar基因在杜氏盐藻巾瞬时表达，而双拷贝碳酸酐酶(DCA1)基因启动子可驱动bar基因在杜氏盐藻中稳定表达。结论：在氦气压力为100L／min条件下微弹轰击2次，可能是基因枪法转化杜氏盐藻的较好转化条件。在转基因杜氏盐藻研究中，DCA1基因启动子可能是一种较为理想的启动子类型。

Transfer of bar gene to Dunaliella salina by microprojectile delivery and detection of transgenic alga

Aim: To investigate the effects of different bombardment conditions and promoters on transformed Dunaliella salina. Method: The bar report gene was introduced into the Dunaliella salina cells by microprojectile delivery system ( Bio-Rad ). The screening culture of transformed algal cells was performed in PKS liquid and solid medium containing 3 mg/L PPT, respectively and the analyses of the transformed Dunaliella salina cells were done. Results: In the 100 L/min pressure from Helium gas, the transformed efficiency of particle bombardment twice was higher than that of once or thrice particle bombardments. The DCA1 gene promoter from Dunaliella salina was able to drive bar gene stable expression, but the CA1 gene promoter was the bar gene transient expression in transformed Dunaliella salina cells. Conclusion: At rupture pressure of 100 L/min, particle bombardment twice using helium-driven biolistics microprojectile delivery system might be a preferable transformation condition. The promoter of DCA1 gene might be an ideal promoter in the research of transgenic Dunaliella salina.