人胎盘生长因子2在毕赤酵母中的表达和纯化

基金项目: 海南省创新能力建设计划省属科研院所技术开发专项资助项目(No.SQ2017JSKF0027)

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

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。
- 胎盘生长因子2 /
- 毕赤酵母 /
- 纯化
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.
- placental growth factor 2(PIGF-2) /
- Pichia pastoris /
- purify

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