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| 牛白细胞介素-2在毕赤酵母中的表达 | |
| 引用本文： | 陈芳,孙红立,曹祥荣,李震,于瑞嵩.牛白细胞介素-2在毕赤酵母中的表达[J].中国兽医学报,2005,25(2):178-182. |
| 作者姓名： | 陈芳 孙红立 曹祥荣 李震 于瑞嵩 |
| 作者单位： | 1. 南京师范大学,生命科学学院,江苏,南京,210097;上海市农业遗传育种重点实验室,上海,201106 2. 南京师范大学,生命科学学院,江苏,南京,210097 3. 上海市农业科学院畜牧兽医研究所,上海,201106;上海市农业遗传育种重点实验室,上海,201106 |
| 基金项目： | 上海市科技兴农重点攻关项目(农科攻字2001第3-7号)资助~~ |
| 摘 要： | 将编码牛白细胞介素-2(BoIL2)成熟肽的cDNA克隆到巴斯德毕赤酵母(Pichia pastoris)表达载体pPICZB中，构建出含BoIL2基因的重组质粒BoIL2-pPICZB。将经Sac Ⅰ酶切后线性化的BoIL2-pPICZB电转化到巴斯德毕赤酵母X-33中，转化子经高浓度Zeoein抗性筛选鉴定后，用1％甲醇诱导目的蛋白表达。经SDS-PAGE及Western blotting检测，表明BoIL2在酵母中获得了胞内表达；通过金属螯合亲和层析(MCAC)获得纯化的重组蛋白；培养小鼠CTLL2细胞进行活性检测，证实所表达的重组BoIL2具有生物活性。 |
| 关 键 词： | 牛 白细胞介素2 毕赤酵母 基因表达 金属螯合亲和层析 |
Expression of Bovine Interleukin-2 Gene in Pichia pastoris |
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| CHEN Fang,SUN Hong-li,CAO Xiang-Rong,LI Zhen,YU Rui-song.Expression of Bovine Interleukin-2 Gene in Pichia pastoris[J].Chinese Journal of Veterinary Science,2005,25(2):178-182. | |
| Authors: | CHEN Fang SUN Hong-li CAO Xiang-Rong LI Zhen YU Rui-song |
| Abstract: | The Interleukin-2 gene cDNA was cloned into the Pichia pastoris expression vector pPICZB,which is under the control of the alcohol oxidase promoter AOX1. The linearized recombinant plasmid of BoIL2-pPICZB,digested by Sac I ,was transformed into X-33 strains by electroporation. The multi-copy insert transformants were screened by Zeocin-resistance and induced by 1% methanol. The intracellular expression products were tested by SDS-PAGE analysis and Western blotting. Purified recombinant BoIL2 was gained by metal-chelating affinity chromatographic (MCAC). Assay with murine CTLL-2 cells showed that the recombinant BoIL2 exhibited the biological activity. |
| Keywords: | bovine Interleukin-2 Pichia pastoris gene expression pPICZB |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |