[Construction and identification of luciferase and EGFP expression vectors containing human IFN-β promoter]

To construct a luciferase expression vector (pGL3-IFNB1) and an enhanced green fluorescent protein expression vector (pGE3-IFNB1) containing human IFN-β promoter, and verify the promoter activity of IFN-β in A549 cells.

IFN-β promoter was amplified with the human genome DNA by PCR, then the segment was cloned into the eukaryotic expression vectors pGL3-basic and pGE3-basic by PCR. The eukaryotic expression vectors were named pGL3-IFNB1 and pGE3-IFNB1, respectively. The recombinant vectors were then transiently transfected into A549 cells respectively, and 6 hours later, the transfected cells were infected with Hantaan virus (HTNV). After 24 hours, the expressions of pGL3-IFNB1 and pGE3-IFNB1 were tested in A549 cells.

Double restriction enzyme digestion and sequence analysis showed that the recombinant vectors were successfully constructed and expressed in A549 cells correctly. After HTNV infection, stronger expression was observed in these cells.

The recombinant vectors pGL3-IFNB1 and pGE3-IFNB1 containing IFN-β promoter have been successfully constructed. It provides useful tools for further study on the mechanism of IFN-β production induced by virus.