| 猪白细胞介素-2的原核表达及其生物学活性研究 |
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| 引用本文： | 吴志明,闫若潜,谢彩华,刘梅芬,刘光辉,黄清.猪白细胞介素-2的原核表达及其生物学活性研究[J].中国畜牧兽医,2009,36(4):89-91. |
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| 作者姓名： | 吴志明 闫若潜 谢彩华 刘梅芬 刘光辉 黄清 |
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| 作者单位： | （河南省动物疫病预防控制中心，郑州 450008） |
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| 摘 要： | 本研究采用RT-PCR方法自刀豆素（Con A）刺激的猪外周血淋巴细胞总RNA中扩增克隆了猪白细胞介素-2（porcine interleutin-2，PoIL-2）成熟肽基因，并亚克隆入pQE30载体进行原核表达，对表达的融合重组猪白细胞介素-2（rPoIL-2）蛋白通过尿素变性、低浓度复性液复性、PBS溶液透析等步骤进行纯化，采用MTT法测定rPoIL-2蛋白促小鼠细胞毒性淋巴样系CTLL-2株细胞增殖活性以评价rPoIL-2的活性。结果表明，成功克隆了rPoIL-2的成熟肽基因，基因全长405 bp，编码134个氨基酸；表达的rPoIFN-α蛋白分子质量大小约为16.5 ku，与预期大小相一致；经纯化后的蛋白质纯度在96%以上；纯化的rPoIL-2蛋白促小鼠CTLL-2株细胞增殖活性最高值是对照细胞的3倍。本研究成功表达了具有生物学活性的PoIL-2蛋白，为新型基因工程抗病毒制剂和免疫增强剂的开发奠定了基础。
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| 关 键 词： | 猪白细胞介素-2 原核表达 纯化 活性 |
Study on the Expression of the Porcine Interleukin-2(PoIL-2) and its Bioactivity |
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| WU Zhi-ming,YAN Ruo-qian,XIE Cai-hua,LIU Mei-Fen,LIU Guang-hui,HUANG Qing.Study on the Expression of the Porcine Interleukin-2(PoIL-2) and its Bioactivity[J].China Animal Husbandry & Veterinary Medicine,2009,36(4):89-91. |
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| Authors: | WU Zhi-ming YAN Ruo-qian XIE Cai-hua LIU Mei-Fen LIU Guang-hui HUANG Qing |
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| Institution: | Henan Centre for Animal Disease Control and Prevention;Zhengzhou 450008;China |
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| Abstract: | The primers were designed to amplify the mature peptide gene of porcine interleukin-2(PoIL-2) gene,and the PCR production was subsequently cloned,sequenced and sub-cloned into the prokaryotic expression vector pQE30.The expressed fusion recombinant PoIL-2(rPoIL-2) protein was purified under the process of denaturing by urea,refolding by a self-innovative renaturation buffer and dialyzing by PBS buffer.The bioactivity of rPoIL-2 was estimated by determining its ability of stimulating the proliferation of IL-... |
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| Keywords: | porcine interleukin-2 prokaryotic expression purify bioactivity |
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