重组人血管内皮生长因子165在Pichia酵母中的表达

目的：采用基因工程方法构建基因工程菌制备人VEGF165蛋白。方法：自人肿瘤组织中获取人VEGF165 cDNA，构建pPIC9K酵母表达载体，采用Pichia酵母真核表达系统表达目的蛋白。结果：SDS-PAGE及Westem-blot结果显示目的蛋白已成功表达。结论：在Pichia酵母中成功表达人VEGF165。

Pichia Expression of Human VEGF165

AIM: A recombinanted DNA Pichia pastroris was constructed to express human vascular endothelial growth factor (hVEGF 165 ) with Pichia pastroris expression system. METHOD:A re comb inant expression plasmid pPIC 9K/hVEGF165 was constructed with pPIC9K vector and human VEGF 165 cDNA prepared from human tumor tissue,then transformed to yeas t host strain GS115 The positive transformants were screened out and cultured in flasks. The supernatant of culture was tested by SDS-PAGE and Western-blot. RESULT:PCR,DNA sequencing and enzyme breakage results showed that the recombined plasmid was constructed successfully. SDS-PAGE and Western-blot re sul t showed that hVEGF was expressed successfully. CONCLUSION: hV EG F 165 was successfully expressed by Pichia pastroris expression system .