人EGFR配体结合区在Pichia酵母中的表达

Pichia Expression of Human EGFR

目的采用基因工程方法构建基因工程菌制备人EGFR配体结合区蛋白。方法 自人肿瘤细胞SK中获取人EGFR cDNA,构建pPIC9K酵母表达栽体,采用Piehia酵母真核表达系统表达目的蛋白。结果SDS-PAGE及Western-blot结果显示目的蛋白已成功表达。结论在Piehia酵母中成功表达人EGFR配体结合区蛋白。

Objective To construct a recombinant DNA Pichia pastroris to express human EGFR ligand combination area with Pichia pastroris expression system. Methods A recombinant expression plasmid pPIC9K/EGFR was constructed with pPIC9K vector and human EGFR cDNA prepared from SK cell, then transformed to yeast host strain GS115. Then the positive transformants were screened out and cultured in flasks. The supernatant of culture was tested by SDS-PAGE and Western-blot. Results PCR,NDNA sequencing and enzyme breakage results showed that the recombined plasmid was constructed successfully. SDS-PAGE and Western-blot result showed that EGFR was expressed successfully. Conclusion EGFR was successfully expressed by Pichia pastroris expression system.