关键词: 人可溶性白细胞介素-1受体, 真核表达载体, 质粒pcDNA 3·1(+)

Abstract:

Objective　The purpose of this study was to construct a eukaryote expression vector carrying human sIL-1R gene.Methods　Both sIL-1R gene and plasmid pcDNA3·1(+) DNAwere digestedwithKpnIandXhoI. After purification, the two fragments obtained were ligated by usingTakaRa DNALigation Kit. This recombinant DNAwas then transformed intoE.coli Competent Cells JM109 and positive cloneswere selected on the LB agarose plate containingAmpicillin (80μg/ml).Results　Six single clones were identified by double digestionwithKpnIandXhoI, and two fragmentswith the size of 5·4 kb and 1·0 kbwere produced as expected.Conclusion　The sIL-1R gene was successfully inserted into the eukaryote expression vector plasmid pcDNA 3·1(+) by the recombination techniquein vitro.

Key words: human soluble interleukin-1 receptor, eukaryote expression vector, plasmid pcDNA 3·1(+)