启动子Pcpcβ提高鱼腥藻7120中hGM-CSF基因表达效率的研究

High Expression of hGM-CSF Induced by the cpcβ Gene Promoter in Anabaena sp.PCC 7120

利用聚球藻7002中编码藻蓝蛋白基因(cpcβ)的启动子(Pcpcβ),替代穿梭表达载体pDC-GM中的启动子PpsbA,启动外源人粒巨噬细胞集落刺激因子(hGM-CSF)基因在鱼腥藻7120中的表达。蛋白免疫印迹证实,在含Pcpcβ启动子的cGM藻株中,hGM-CSF基因的表达水平比原先构建的含PpsbA启动子的GM藻株中提高了90%,且该基因的高效表达对宿主细胞的生长未产生明显影响。结果表明,对于在蓝藻中表达hGM-CSF基因而言,Pcpcβ启动子可能是较适宜的强启动子之一。

To increase the expression amount of foreign hGM-CSF in Anabaena sp. PCC 7120, the PpsbA promoter was replaced by the cpcβ gene promoter, originating from Synechococcus sp. PCC 7002. Western blot analysis showed that the expression level of hGM-CSF induced by the cpcβ gene promoter was about 90 % more than that induced by the PpsbA promoter. Thus, the cpcβ gene promoter is a highly efficient promoter, and remarkably stimulates over-expression of hGM-CSF in Anabaena sp. PCC 7120.