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Updated: Aug 17, 2026
Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
Published on: February 9, 2015
[Construction of eukaryote expression vector carrying human soluble interleukin-1 receptor gene]
Peng Li1, Yan Xu, Yunhui Zhang
1Department of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, Chengdu 610041, China.
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 pcDNA 3.1(+) DNA were digested with KpnI and XhoI. After purification, the two fragments obtained were ligated by using TakaRa DNA Ligation Kit. This recombinant DNA was then transformed into E. coli Competent Cells JM109 and positive clones were selected on the LB agarose plate containing Ampicillin (80 micrograms/ml).
Results:
Six single clones were identified by double digestion with KpnI and XhoI, and two fragments with the size of 5.4 kb and 1.0 kb were produced as expected.
Conclusion:
The sIL-1R gene was successfully inserted into the eukaryote expression vector plasmid pcDNA 3.1(+) by the recombination technique in vitro.
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