| 鸡IL-2的毕赤酵母表达和生物活性鉴定 |
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| 引用本文： | 张鹏 王黎霞 安,健.鸡IL-2的毕赤酵母表达和生物活性鉴定[J].中国农学通报,2010,26(18):1-5. |
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| 作者姓名： | 张鹏 王黎霞 安 健 |
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| 作者单位： | 1. 北京农学院动物科技系,北京,102206
2. 北京农业职业学院畜牧兽医系,北京,102442 |
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| 基金项目： | 北京市教委资助项目"柔嫩艾美耳球虫免疫细胞因子佐剂研究"，北京市属市管高校人才强教计划项目"中青年骨干教师计划" |
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| 摘 要： | IL-2是一种重要的免疫相关细胞因子，借助PCR去掉鸡IL-2基因片段(387bp)信号肽，连接到pPICZαA表达载体中，通过电转化转入Pichia酵母GS115菌株的感受态中，对筛选出来的阳性菌株以终浓度为1%的甲醇诱导表达，经SDS-PAGE及Western Blot分析，结果表明酵母成功分泌表达了重组鸡IL-2，分子量为17.8kD。通过Ni-NTA His Bind 柱层析，250mM咪唑的洗脱液洗脱，得到纯度较高的重组鸡IL-2，淋巴细胞增殖实验证明，重组鸡IL-2具有明显的促进淋巴细胞增殖的生物活性，为进一步研究其免疫佐剂的作用提供了实验基础。
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| 关 键 词： | 水稻 水稻 锌指蛋白 基因克隆 表达 |
| 收稿时间： | 2010/3/31 0:00:00 |
| 修稿时间： | 2010/4/29 0:00:00 |
Expression in Pichia pastoris and biological activity characterization of recombinant chicken IL-2 |
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| Zhang Peng,Wang Lixia,An Jian.Expression in Pichia pastoris and biological activity characterization of recombinant chicken IL-2[J].Chinese Agricultural Science Bulletin,2010,26(18):1-5. |
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| Authors: | Zhang Peng Wang Lixia An Jian |
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| Institution: | Zhang Peng1,Wang Lixia2,An Jian1 (1 Beijing Agriculture University,Beijing 102206,2Beijing Vocational College of Agriculture,Beijing 102442) |
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| Abstract: | Chicken IL-2 is an key cytokine for immunity. Removed the signal peptide by PCR，chIL-2 gene fragments (387bp) was connected to the pPICZαA expression vector and transformed to Pichia pastoris GS115 by electroporation. SDS-PAGE and Western Blot analysis showed that chIL-2(17.8kD) was successfully expressed and secreted by positive strains induced by 1% methanol. The chIL-2, which was purified by Ni-NTA His Bind column chromatography with 250mM imidazole elution buffer, had significant biological activity to promote lymphocyte proliferation, which provides the experimental basis for the further research on chIL-2 as a kind of immune adjuvant. |
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