[The cloning and expression of human secreted interleukin 2 receptor alpha chain gene in eukaryotic cells].
By deleting the sequences encoding the cytoplasmic domain, transmembrane domain and the 5' untranslated region, human interleukin-2 receptor alpha cDNA fragment was subcloned into an eukaryotic expression vector pRc/CMV. The recombinant plasmid was then transfected into CHO cells by either electroporation or calcium phosphate precipitation. 11 G418-resistent clones were randomly selected for detection of mRNA expression. 5 clones out of the 11 were proved to be positively hybridyzed with IL-2R alpha cDNA probe. All of the 5 positive cell lines also expressed sIL-2R alpha in their supernatants as detected by ELISA. The establishment of rhsIL-2R alpha cell lines is of importance in the detection and purification of IL-2 based on the ability of affinity binding between IL-2 and its recombinant receptor.
Cheng
X
X
Institute of Basic Medical Sciences, CAMS and PUMC, Beijing.
Du
K
K
Heng
F
F
Wu
N
N
Shen
Y
Y
D000818:Animals; D016466:CHO Cells / Q000378:metabolism*; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004274:DNA, Recombinant / Q000096:biosynthesis; D005057:Eukaryotic Cells / Q000378:metabolism; D015870:Gene Expression; D010957:Plasmids; D012333:RNA, Messenger / Q000096:biosynthesis; D015375:Receptors, Interleukin-2 / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D004274:DNA, Recombinant; D012333:RNA, Messenger; D015375:Receptors, Interleukin-2