Expression vector as well as construction method and application thereof

The invention relates to an expression vector for gene expression in bacteria and mammalian cells as well as a construction method and application of the expression vector. The expression vector comprises a bacterial expression cassette and a mammal expression cassette, wherein the bacterial expression cassette and the mammal expression cassette comprise a CMV (cytomegalovirus) promoter, an L-rhamnose induced rhaB promoter, a Shine-Dalgarno (SD) sequence, a target fluorescent protein gene, a bacterial rrnB T1 terminator and an SV40poly-A tail; and the reference gene expression box comprises an SV40 promoter, a reference FP gene and an HSV-TK poly (A) tail. The method combines the speed and convenience of high throughput screening of large bacterial libraries with mammalian cell-based high content screening for intracellular brightness and photostability, without the need for a re-cloning step. 本发明涉及一种用于细菌和哺乳动物细胞中的基因表达的表达载体及其构建方法和应用。所述表达载体包括：细菌和哺乳动物表达盒，其包括CMV启动子、L-鼠李糖诱导的rhaB启动子、Shine-Dalgarno(SD)序列、靶荧光蛋白基因、细菌r