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梁小庆, 闫鹏飞, 刘煜. 人胎盘生长因子2在毕赤酵母中的表达和纯化[J]. 中国药科大学学报, 2018, 49(3): 376-380. DOI: 10.11665/j.issn.1000-5048.20180318
引用本文: 梁小庆, 闫鹏飞, 刘煜. 人胎盘生长因子2在毕赤酵母中的表达和纯化[J]. 中国药科大学学报, 2018, 49(3): 376-380. DOI: 10.11665/j.issn.1000-5048.20180318
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LIANG Xiaoqing, YAN Pengfei, LIU Yu. Expression and purification of human placental growth factor 2 using the Pichia pastoris expression system[J]. Journal of China Pharmaceutical University, 2018, 49(3): 376-380. DOI: 10.11665/j.issn.1000-5048.20180318
Citation: LIANG Xiaoqing, YAN Pengfei, LIU Yu. Expression and purification of human placental growth factor 2 using the Pichia pastoris expression system[J]. Journal of China Pharmaceutical University, 2018, 49(3): 376-380. DOI: 10.11665/j.issn.1000-5048.20180318
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人胎盘生长因子2在毕赤酵母中的表达和纯化

Expression and purification of human placental growth factor 2 using the Pichia pastoris expression system

    摘要
  • 摘要: 为获得真核表达的重组人胎盘生长因子2(PIGF-2)纯化蛋白样品，采用基因工程方法构建重组人PIGF-2酵母表达载体pPIC9K-PIGF-2，电转后筛选阳性克隆，经甲醇诱导表达、肝素亲和柱色谱分离纯化得到蛋白纯化样品，经SDS-PAGE和Western blot鉴定，该蛋白为重组人PIGF-2纯化蛋白，表明该法可用于制备分泌表达的hPIGR-2。

     

    Abstract: To prepare the human placental growth factor 2(PIGF-2), human PIGF-2 gene was cloned into pPIC9K vector to construct the recombinant pPIC9K-PIGF-2 vector. Linearized recombinant pPIC9K-PIGF-2 was transformed into Pichia pastoris by electroporation. YPD-Geneticin plate was used to screen geneticin hyper-resistant colonies. The positive colonies were verified by PCR. Results of SDS-PAGE and Western blot showed that recombinant human PIGF-2 was expressed after being induced by methanol. Using its characteristic heparin binding, recombinant human PIGF-2 was successfully purified by heparin affinity column chromatography.

     

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