摘要
目的 构建表达有特异细胞毒性融合蛋白的工程质粒。方法 利用PCR扩增出PE3 5KDEL基因片段 ,经酶切后插入EGF PET2 8A载体 ,并转化至BL2 1(DE3 )中。采用Westernblot对重组毒素进行鉴定。结果EGF PE3 5KDEL重组毒素占菌体总蛋白的 2 0 % ,分子量约为 410 0 0。结论 EGF PE3
Objective To construct a engineering plasmid which can express special cytotoxic fusion protein.Methods Amplify PE35KDEL DNA fragment by PCR,digest with restriction endonuclease,insert into EGF PET28a vector and transform to BL21(DE3) for expression under induction of IPTG.The expressed product was identified by Western blot.Results The expressed EGF PE35KDEL toxin contained 20% of total somatic protein,and the relative molecular weight of it was about 41 000.Conclusion The successful construction and expression of EGF PE35KDEL laid a foundation for developing potential targeting antitumor medicine.
出处
《中国生物制品学杂志》
CAS
CSCD
2003年第2期68-70,共3页
Chinese Journal of Biologicals
