Iwata H, Yamamoto M, Hasegawa A, Kurata K, Inoue T
Department of Veterinary Hygiene, Faculty ofAgriculture, Yamaguchi University, Japan.
J Vet Med Sci. 2000 Oct;62(10):1101-4. doi: 10.1292/jvms.62.1101.
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.
使用pGEX载体在大肠杆菌中表达了一种不含信号肽的成熟形式的猪白细胞介素-2（IL-2）蛋白，作为谷胱甘肽S-转移酶（GST）融合蛋白。由于在SDS-PAGE分析中，大部分GST-IL-2融合蛋白在不溶性部分被检测到，因此使用尿素通过重折叠程序使不溶性融合蛋白溶解。重组IL-2（rIL-2）通过使用谷胱甘肽琼脂糖4B的批量方法和因子Xa消化进行纯化，并用于在小鼠中制备抗血清。在免疫印迹分析中，抗血清与在杆状病毒系统中表达的rIL-2发生反应。此外，纯化的rIL-2在CTLL-2增殖反应中显示出高生物活性。
