Construction of eukaryote expression vector carrying human soluble interleukin-1 receptor gene.
- Author:
Peng LI
1
;
Yan XU
;
Yunhui ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Cloning, Molecular; DNA, Recombinant; genetics; Eukaryotic Cells; metabolism; Genetic Vectors; Humans; Plasmids; genetics; immunology; Polymerase Chain Reaction; RNA, Messenger; genetics; Receptors, Interleukin-1; biosynthesis; genetics; Recombinant Proteins; biosynthesis; genetics; Recombination, Genetic
- From: West China Journal of Stomatology 2003;21(2):140-143
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEThe purpose of this study was to construct a eukaryote expression vector carrying human sIL-1R gene.
METHODSBoth 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).
RESULTSSix 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.
CONCLUSIONThe sIL-1R gene was successfully inserted into the eukaryote expression vector plasmid pcDNA 3.1(+) by the recombination technique in vitro.
