| 猪IL-18 在杆状病毒/昆虫细胞中的表达 |
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| 引用本文： | 王振亚,王彦彬,陈红英,邵攀峰,宁晓东,潘娜,韩利静,崔保安.猪IL-18 在杆状病毒/昆虫细胞中的表达[J].生物工程学报,2011,27(1):118-123. |
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| 作者姓名： | 王振亚 王彦彬 陈红英 邵攀峰 宁晓东 潘娜 韩利静 崔保安 |
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| 作者单位： | 1. 河南农业大学牧医工程学院,郑州,4500021;河南省动物件食品安全重点实验室,郑州,450002
2. 河南农业大学牧医工程学院,郑州,4500021 |
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| 基金项目： | 国家“十一五”科技支撑计划 (No. 2006BAD06A08) 资助。 |
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| 摘 要： | IL-18作为一种多效性细胞因子，在机体免疫应答及各种生理功能中发挥着重要的调节作用，根据猪IL-18基因序列设计1对引物，将编码猪IL-18的成熟蛋白基因亚克隆到杆状病毒转移载体pFastBacDual中，并在C端融合6个组氨酸标签以利于纯化，然后转化到含穿梭载体Bacmid的感受态细胞DH10Bac中，发生转座作用。将重组质粒转染昆虫细胞，SDS-PAGE可检测到分子量为18 kDa左右的重组蛋白，Western blotting证实该重组蛋白可与兔抗猪IL-18抗体发生特异性反应。纯化后蛋白能明显促进猪T淋巴细胞转化，表明所表达的IL-18具有较高的生物活性。此研究为进一步开发研制新型免疫佐剂奠定了基础。
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| 关 键 词： | 猪白细胞介素-18，昆虫细胞，表达 |
| 收稿时间： | 2010/4/12 0:00:00 |
| 修稿时间： | 2010/7/19 0:00:00 |
Expression of porcine Interleukin-18 in baculovirus/insect cells |
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| Zhenya Wang,Yanbin Wang,Hongying Chen,Panfeng Shao,Xiaodong Ning,Na Pan,Lijing Han and Baoan Cui.Expression of porcine Interleukin-18 in baculovirus/insect cells[J].Chinese Journal of Biotechnology,2011,27(1):118-123. |
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| Authors: | Zhenya Wang Yanbin Wang Hongying Chen Panfeng Shao Xiaodong Ning Na Pan Lijing Han and Baoan Cui |
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| Institution: | College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; Henan Key Laboratory of Animal-DERIVED Food Safety of Henan Province, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; Henan Key Laboratory of Animal-DERIVED Food Safety of Henan Province, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; Henan Key Laboratory of Animal-DERIVED Food Safety of Henan Province, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; Henan Key Laboratory of Animal-DERIVED Food Safety of Henan Province, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;College of Animal Husbandry and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China; Henan Key Laboratory of Animal-DERIVED Food Safety of Henan Province, Zhengzhou 450002, China |
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| Abstract: | 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 6×Histidine 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. |
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| Keywords: | porcine Interleukin-18 insect cells expression |
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