重组植物AHA2蛋白的真核表达及纯化研究

Eukaryotic Expression and Purification of Recombinant Plant AHA2 Protein

摘要:

AHA2蛋白位于植物细胞细胞质质膜上，该蛋白在原核蛋白表达体系中无法重组表达.已经报道的酿酒酵母（Saccharomyces cerevisiae）表达体系操作复杂、成本高昂.本文利用RT-PCR、分子克隆等技术构建了植物AHA2蛋白的真核表达载体，并利用毕赤酵母（PiChia）对AHA2蛋白加以表达.利用亲和层析和分子筛层析相结合的方法，获得了高质量的处于高活性态的目的蛋白.

Abstract:

AHA2 anchors within the cytoplasm membrane of plant cell. In prokaryotic expression system, the protein can’t be recombinant expressed. In Saccharomyces cerevisiae expression system, the procedures of AHA2 purification stay complex and expensive. By means of RT-PCR and molecular cloning, we constructed an eukaryotic expression vector of AHA2 with PiChia as host. By method of affinity chromatography and gel filtration chromatography, we obtained target protein in highly active with high quality.