菜热休克蛋白基因HSP81.1启动子的克隆与功能验证

本研究从甘蓝型油菜（Brassica napus）基因组中扩增获得热休克蛋白基因启动子,并将其克隆至双元载体pBI121的GUS报告基因上游,构成双元表达载体pBI121 HSP GUS。通过农杆菌（Agrobacterium tumefaciens）介导法转化烟草后获得转基因植株。对T1转基因植株研究表明,外源基因在42℃处理1h后开始表达,且即使用同样条件诱导,GUS基因的表达率和表达量在不同生长时期也有差异。在第50d左右的苗期,GUS基因的表达率和表达量高,但随植株老化而逐渐下降。研究显示,利用油菜的热休克蛋白启动子可以达到通过温度控制烟草基因表达的目的。

Cloning and function analysis of promoter of heat shock protein 81.1 from Brassica napus L.

Promoter of temperature-induced heat shock protein（HSP） gene is used to control the exogenous gene expression not only to improve the expression accuracy,but also to enhance the safety of GM（genetically modified） crop.In present study,a HSP gene promoter from Brassica napus L.was cloned onto the upstream of GUS reporter gene in binary vector pBI121,in order to construct a binary expression vector pBI121 HSP GUS.The vector was then transformed into tobacco by Agrobacterium tumefaciens-mediated method.It was shown that T1 plants started to express the foreign GUS gene at 42℃ induction for 1h.The expression rate and amount of GUS gene varied at different growth stages and reached to a high level around the 50th day after seeding.Then the expression decreased gradually.The results indicated a potential of HSP promoter from rapeseed to control the expression of tobacco target gene under heat condition.