title: [Expression of goose interleukin-2 gene in Escherichia coli and isolation of its soluble monomer].
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.
authors.affiliation: Laboratory of Virology and Immunology, Institute of Preventive Veterinary Medicine, Zhejiang University, Hangzhou 310029, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Qi
authors.forename: Jigang
authors.initials: J
authors.lastname: Chen
authors.forename: Jinyong
authors.initials: J
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Fang
authors.forename: Jiajun
authors.initials: J
authors.lastname: Wu
authors.forename: Jiyong
authors.initials: J
authors.lastname: Zhou
mesh_terms: D000818:Animals; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005777:Geese / Q000235:genetics*; D002479:Inclusion Bodies / Q000378:metabolism; D007376:Interleukin-2 / Q000096:biosynthesis / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012995:Solubility
chemical_list: D007376:Interleukin-2; D011994:Recombinant Proteins