Construction and expression of the overexpression vector of steroid xenobiotic receptor gene Nr1i2 in rat liver

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