Expression of goose interleukin-2 gene in Escherichia coli and isolation of its soluble monomer.
- Author:
Jing QI
1
;
Jigang CHEN
;
Jinyong WANG
;
Jie FANG
;
Jiajun WU
;
Jiyong ZHOU
Author Information
1. Laboratory of Virology and Immunology, Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Escherichia coli;
genetics;
metabolism;
Geese;
genetics;
Inclusion Bodies;
metabolism;
Interleukin-2;
biosynthesis;
genetics;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification;
Solubility
- From:
Chinese Journal of Biotechnology
2008;24(2):183-187
- CountryChina
- Language:Chinese
-
Abstract:
Recombinant expression plasmid of pET-28a (+)-goIL-2 was constructed by inserting the goose IL-2 gene without the signal peptide sequence into the prokaryotic expression vector pET-28a (+), and transformed into the bacterial competent E. coli BL21 (DE3) cells for expression. After IPTG induction, an expected protein band with molecular weight of 15.0 kD was observed on SDS-PAGE gel, recognized by monoclonal antibody against goose IL-2 in western-blotting assay. In the pET-28a (+) expression system, much of the recombinant goose IL-2 (rgoIL-2) was found in inclusion bodies with a portion of soluble protein. The monomer and multimers of soluble goose interleukin 2 proteins were observed in native electrophoresis. The rgoIL-2 proteins were purified by Ni-NTA column under a native condition. The rgoIL-2 soluble protein monomer was isolated by a quick protein isolation and purification system of AKTA FPLC and identified by native PAGE. Bioactivity analysis showed that the rgoIL-2 monomer stimulated the proliferation of goose lymphocytes in vitro. This will establish a basis for further study about the biological function and clinical application of goose IL-2.
