Cloning, expression and purification of the ligand-binding region of human IL-6R in E. coli and its preliminary functional identification.
The ligand-binding region of human IL-6R is taken as the target gene fragment to be cloned and expressed. With pET-3b as expressing vector, two recombinants pET-6R(B) and pET-6R(B)4 have been constructed encoding the ligand-binding region (28 kD) of hIL-6R and its dimmer (53 kD), respectively. After induction with IPTG, they produced two proteins rIL6R-28 of 28 kD and rIL6R-53 of 53 kD amounting to 50% and 30% of total bacteria proteins, respectively. The expressed products were mainly recovered as inclusion bodies. After purification and renaturation, both of them were capable of augmenting the growth-stimulating effect of IL-6 on 7TD1 cells, an IL-6 dependent cell line. The result of ELISA also revealed that both rIL6R-28 and rIL6R-53 had the obvious ligand-binding activity.
Duan
J
J
Institute of Basic Medical Sciences, Beijing, China.
Wang
J
J
Han
J
J
Peng
S
S
Zou
M
M
Miao
J
J
Zhao
C
C
Ma
X
X
D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D018123:Receptors, Interleukin / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D019947:Receptors, Interleukin-6
D015703:Antigens, CD; D004274:DNA, Recombinant; D015850:Interleukin-6; D010446:Peptide Fragments; D018123:Receptors, Interleukin; D019947:Receptors, Interleukin-6