[Construction and expression of eukaryotic expression vector pBK-IL-1ra].
OBJECTIVE
Construction of eukaryotic vector with high expression of interleukin-1 receptor antagonist (IL-1ra) for IL-1ra gene delivery.


METHODS
(1) Eukaryotic expression vector pBK-IL-1ra was constructed by recombinant DNA technique, and identified by restriction mapping enzymes, PCR, and DNA sequence analysis. (2) Expressions of the IL-1ra mRNA and IL-1ra in vitro and in vivo were detected by in situ hybridization, ELISA, and immunohistochemistry.


RESULTS
(1) The correct construction of pBK-IL-1ra was identified by the method above. (2) The higher levels of IL-1ra in the supernate of COS-7 cells (24 h, (P < 0.001; 48 h, P < 0.01) and the over expression of IL-1ra mRNA (P < 0.01) and IL-1ra (P < 0.05) in the synoviocytes have been detected after 48 h of the transfection with pBK-IL-1ra. (3) The over expression of IL-1ra in muscle of the mice was detected by immunohistochemistry after 4 days of pBK-IL-1ra injection (P < 0.01)


CONCLUSIONS
We have successfully constructed eukaryotic expression vector pBK-IL-1ra which could highly expressed IL-1ra in vitro and in vivo, and it offers a novel possibility in the research of IL-1ra gene therapy.
Yang
G
G
Department of Rheumatology, PUMC Hospital, CAMS, PUMC, Beijing 100730, China.
Li
F
F
Jiang
M
M
Di
C
C
Huang
J
J
D000818:Animals; D019556:COS Cells; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D053590:Interleukin 1 Receptor Antagonist Protein; D051379:Mice; D008811:Mice, Inbred DBA; D010957:Plasmids / Q000235:genetics*; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000235:genetics*; D020298:U937 Cells
C507763:IL1RN protein, human; C507764:Il1rn protein, mouse; D053590:Interleukin 1 Receptor Antagonist Protein; D012795:Sialoglycoproteins