Abstract
Objective To construct the overexpression plasmid of steroid xenobiotic receptor (SXR) gene Nrli2, and verify its expression by HEK-293 cells. Methods The inverse transcription of total RNA in rat liver was amplified to Nrli2 coding frame with enzyme loci primer, and the Nrli2 was sub- cloned into the pcDNA3.1, which was subjected to enzyme digestion and verified by sequencing. pcDNA3.1 ( + ) -Nrl i2 was transfected into 293 cells, and 48 h later, the cells were harvested for fluores- cent quantitative polymerase chain reaction ( FQ-PCR), and 72 h later for Western blotting. Results The Nrl i2 coding region was successfully amplified, and cloned into the vector pcDNA3.1. The expression of SXR gene Nrli2 in HEK-293 cells and pcDNA-3. 1 ( + ) transfected group was significantly lower than in pcDNA3.1 ( + ) -Nrl i2 transfected group ( both P 〈 0. 01 ). Conclusion We have successfully constructed Nrli2 eukaryotic overexpression vector, which was successfully expressed in 293 cells. Key words: Steroid; Xenobiotic receptor; Vector construction ; Gene cloning
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