Expression of biologically active recombinant ferret (Mustela putorius furo) interleukin-8 from Escherichia coli.
The authors expressed recombinant ferret interleukin-8 protein (rfrIL-8) in Escherichia coli as a glutathione-S-transferase fusion protein. Western blot analyses revealed that anti-ovine IL-8 antibody reacted with rfrIL-8 at 10 kDa. To confirm that the rfrIL-8 was biologically active, the authors examined chemotaxis and respiratory burst activity of ferret polymorphonuclear blood cells (PMNs) exposed to rfrIL-8. The rfrIL-8 strongly induced chemotactic and respiratory burst activities in a statistically significant manner as compared with a negative control. Thus, the authors were able to successfully express biologically active rfrIL-8.
Nakata
Makoto
M
Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan.
Kozue
Yu
Y
Itou
Takuya
T
Sakai
Takeo
T
D000818:Animals; D000918:Antibody Specificity; D001483:Base Sequence; D015153:Blotting, Western; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005289:Ferrets / Q000235:genetics* / Q000276:immunology*; D015870:Gene Expression; D007120:Immunochemistry; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics* / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D016897:Respiratory Burst / Q000187:drug effects; D012756:Sheep
D017931:DNA Primers; D016209:Interleukin-8; D011993:Recombinant Fusion Proteins