[Cloning, expression and identification of human interleukin-10 gene].
OBJECTIVE
To construct a prokaryotic expression vector containing human interleukin-10 (IL-10) gene, express and identify the protein.


METHODS
Human IL-10 gene from HepG2 was isolated and identified by DNA sequencing, and then cloned into the expression vector PET-28 a (+) to construct prokaryotic expression vector. The recombinant protein was expressed in BL21(DE3) and identified by Western blot and ELISA.


RESULTS
The expressed protein was mainly located in the inclusion body. The relative molecular mass of the expressed product was identical to that of prediction. The expressed protein had binding activity with specific antibody.


CONCLUSIONS
The human IL-10 gene was successfully cloned and expressed in E.coli. The expressed product had antigenicity which provides foundation for preparation of monoclonal antibodies against IL-10.
Sun
Jie
J
Department of Immunology, Beijing Institute of Infections Diseases, Beijing, China. energysj@yeah.net
Li
Bo-An
BA
Qi
Yang
Y
Zhou
Ji-Wen
JW
Zhu
Lei
L
Chi
Shu-Ping
SP
Cheng
Yun
Y
D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D016753:Interleukin-10 / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding
D016753:Interleukin-10