[Construction of eukaryote expression vector carrying human soluble interleukin-1 receptor gene].
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.


CONCLUSIONS
The sIL-1R gene was successfully inserted into the eukaryote expression vector plasmid pcDNA 3.1(+) by the recombination technique in vitro.
Li
Peng
P
Department of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, Chengdu 610041, China.
Xu
Yan
Y
Zhang
Yunhui
Y
D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics*; D005057:Eukaryotic Cells / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D011995:Recombination, Genetic
D004274:DNA, Recombinant; D012333:RNA, Messenger; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins