Wang Zhenya, Wang Yanbin, Chen Hongying, Shao Panfeng, Ning Xiaodong, Pan Na, Han Lijing, Cui Baoan
College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China.
Sheng Wu Gong Cheng Xue Bao. 2011 Jan;27(1):118-23.
IL-18, as a polyphonic cytokine, is important in immune response and physiologic function. We designed one pair of primers, amplified the porcine IL-18 gene fused with a C-terminal 6xHistidine tag, and then subcloned into the pFastBacDual of Baculovirus transfer vector and transformed into DH10Bac containing a shuttle vector of Bacmid. After co-transfecting the recombinant plasmid into insect cells, the 18 kDa expressed protein of porcine IL-18 was detected by SDS-PAGE; the specificity of expressed protein was confirmed by Western blotting. The purified porcine IL-18 protein induced obvious proliferation of porcine T lymphocytes in vitro, which indicated that the expression of IL-18 had high biological activity.
白细胞介素-18(IL-18)作为一种多效性细胞因子,在免疫应答和生理功能中发挥着重要作用。我们设计了一对引物,扩增出与C末端6x组氨酸标签融合的猪IL-18基因,随后亚克隆至杆状病毒转移载体pFastBacDual中,并转化至含有杆粒穿梭载体的DH10Bac中。将重组质粒共转染至昆虫细胞后,通过SDS-PAGE检测到猪IL-18的18 kDa表达蛋白;通过蛋白质免疫印迹法(Western blotting)证实了表达蛋白的特异性。纯化后的猪IL-18蛋白在体外诱导猪T淋巴细胞明显增殖,这表明IL-18的表达具有高生物学活性。