Nakata Makoto, Kozue Yu, Itou Takuya, Sakai Takeo
Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan.
Vet Immunol Immunopathol. 2010 Nov 15;138(1-2):114-7. doi: 10.1016/j.vetimm.2010.06.017. Epub 2010 Jul 6.
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.
作者在大肠杆菌中将重组雪貂白细胞介素-8蛋白（rfrIL-8）表达为谷胱甘肽-S-转移酶融合蛋白。蛋白质印迹分析显示，抗羊IL-8抗体与10 kDa的rfrIL-8发生反应。为了确认rfrIL-8具有生物活性，作者检测了暴露于rfrIL-8的雪貂多形核血细胞（PMN）的趋化性和呼吸爆发活性。与阴性对照相比，rfrIL-8以统计学上显著的方式强烈诱导趋化性和呼吸爆发活性。因此，作者能够成功表达具有生物活性的rfrIL-8。
