[Prokaryotic expression, purification and biological activity of recombinant human IL-17/His protein].
OBJECTIVE
To investigate the biological activity of recombinant human IL-17 protein in vitro.


METHODS
The gene region of human IL-17 was cloned by RT-PCR. After identification by sequencing, the hIL-17 gene encoding function domain was cloned into expression plasmid PQE3.0 and transfect into E.coli M15. By the induction of Isopropyl-beta-D-Thiogalacto-Pyranoside (IPTG), recombinant IL-17/His protein was effectively expressed in E.coli M15. The recombinant protein was identified by Western blot.


RESULTS
After denaturation, renaturation and purification by HiTrap affinity column, the recombinant protein stimulated HeLa, a human uterine cervix cancer cell line, to excrete IL-6 and GM-CSF in vitro.


CONCLUSIONS
IL-17/His recombinant protein is of high biological activity, which can be used to make further study of auto-immune diseases.
Liu
Gao-qin
GQ
Institute of Medical Biotechnology, Soochow University, Suzhou, China. liugaoqin2006@sina.com
Wu
Hong-ya
HY
Zhang
Guang-bo
GB
Ma
Hong-bing
HB
Ma
Zhen-ni
ZN
Ju
Song-guang
SG
Zhang
Xue-guang
XG
D015153:Blotting, Western; D002478:Cells, Cultured; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D020381:Interleukin-17 / Q000235:genetics* / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D020381:Interleukin-17; D015850:Interleukin-6; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor