欢迎访问《中国农学通报》，
中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (18): 1-5.

所属专题： 畜牧兽医

• 畜牧 动物医学 蚕 蜂 •    下一篇

鸡IL-2的毕赤酵母表达和生物活性鉴定

张鹏 王黎霞 安 健   

  • 收稿日期:2010-03-31 修回日期:2010-04-29 出版日期:2011-09-20 发布日期:2011-09-20
  • 基金资助:
    北京市教委资助项目; 北京市属市管高校人才强教计划项目

Expression in Pichia pastoris and biological activity characterization of recombinant chicken IL-2

  • Received:2010-03-31 Revised:2010-04-29 Online:2011-09-20 Published:2011-09-20

PDF (PC)

1381

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。

关键词: 水稻, 水稻, 锌指蛋白, 基因克隆, 表达

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.