摘要
目的 :采用基因工程方法构建基因工程菌制备人VEGF1 65蛋白。方法 :自人肿瘤组织中获取人VEGF1 65cDNA ,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Western 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VEGF1 65。
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 .
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2003年第6期577-581,共5页
Journal of China Pharmaceutical University
基金
上海市科技发展基金(No .0 2 4 31 91 0 5)
