关键词: 质粒, SDF2L1, 基因, 表达

Abstract: Objective To determine the expression levels of SDF2L1 in mouse liver under different physiological and pathophysiological conditions and to perform SDF2L1 overexpression by using eukaryotic expression system for further functional research. Methods The expression levels of SDF2L1 in mouse liver under different conditions were determined by Real-time PCR. Primers were designed commercially. Total RNA was isolated from the wild type C57BL/6J mouse liver and reverse transcribed to cDNA. The coding sequence of SDF2L1 was amplified by PCR, using the cDNA as template. The product of PCR reaction was purified and inserted into pcDNA4/myc-His vector and then transformed into E.coli competent cells. Expression of recombinant was transfected into 293A cells and identified by SDS-PAGE. Result The fasting or pathological condition lead to decreased expression levels of SDF2L1. The molecular weight of purified plasmid was 5.8kb. The product was 666bp and its sequence was identical to that in wide type SDF2L1. The molecular of the recombinant protein was 26KD. Conclusion Our data suggest SDF2L1 may play a significant role in the regulation of hepatic glucose and lipid metabolism.

Key words: plasmid, SDF2L1, gene, expression