[Cloning of hIL-1Ra gene and its expression in E.coli].
OBJECTIVE
To clone human IL-1Ra gene and express it in E.coli.


METHODS
Human IL-1Ra cDNA was obtained by RT-PCR with the the total RNA extracted from human peripheral blood leucocytes as template. The cDNA was cloned into pBV220 vector and expressed. The expressed product was renatured and purified. Separation, purification and bioactivity analysis of the expressed products were performed.


RESULTS
IL-1Ra gene was successfully expressed in E.coli and the expression level reached to about 40% of total bacteria protein. The purity of the final product was over 98%. The product could obviously suppress the secretion of IL-2 by EL-4 cells stimulated with IL-1beta.


CONCLUSIONS
The expression of hIL-1Ra gene with bioactivity in E.coli lays experimental foundation for further development and utilisation.
Chang
Shao-hong
SH
Insititute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China.
Wu
Jun
J
Gong
Xin
X
D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D002479:Inclusion Bodies / Q000378:metabolism; D053590:Interleukin 1 Receptor Antagonist Protein; D007376:Interleukin-2 / Q000378:metabolism; D051379:Mice; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D012795:Sialoglycoproteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D014407:Tumor Cells, Cultured
C507763:IL1RN protein, human; C507764:Il1rn protein, mouse; D053590:Interleukin 1 Receptor Antagonist Protein; D007376:Interleukin-2; D011994:Recombinant Proteins; D012795:Sialoglycoproteins