Expression of porcine interleukin-2 in Escherichia coli.
A mature form of porcine interleukin-2 (IL-2) protein without signal peptides was expressed as glutathione S-transferase (GST) fusion proteins in Escherichia coli using pGEX vector. Since most of GST-IL-2 fusion protein was detected in an insoluble fraction on SDS-PAGE analysis, the insoluble fusion protein was solubilized by refolding procedure using urea. The recombinant IL-2 (rIL-2) was purified by a batch method using Glutathione Sepharose 4B and factor Xa digestion and used for preparation of antisera in mice. The antisera reacted with rIL-2 expressed in baculovirus system on immunoblot analysis. In addition, the purified rIL-2 showed a high biological activity on CTLL-2 proliferative response.
Iwata
H
H
Department of Veterinary Hygiene, Faculty ofAgriculture, Yamaguchi University, Japan.
Yamamoto
M
M
Hasegawa
A
A
Kurata
K
K
Inoue
T
T
D000818:Animals; D002846:Chromatography, Affinity / Q000662:veterinary; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004926:Escherichia coli / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005822:Genetic Vectors; D007376:Interleukin-2 / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013552:Swine / Q000235:genetics / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic / Q000378:metabolism
D007376:Interleukin-2; D011993:Recombinant Fusion Proteins