摘要
利用构建好的质粒pPIC9K-RGD-4C-His Tag-AGAP(analgesic-antitumor peptide,抗癌镇痛肽)进行单酶切线性化,然后电转化毕赤酵母GS115,电转之后的菌落通过菌落PCR进行筛选,最终得到阳性菌落。阳性菌落经增菌培养,甲醇诱导后的上清SDS-PAGE电泳后经western blot检测证明,抗癌镇痛肽肺靶向融合体已经成功得到表达。
A constructed plasmid pPIC9K-RGD-4C-His Tag-AGAP（analgesic-antitumor peptide） was linearized with single enzyme digestion and electroporation transformed into Pichia pastoris GS115,the transformed colonies were screened through colony PCR and finally obtained positive ones.And they were proliferated,and the supernatant SDS-PAGE after induced with methanol and tested proved with western blot that fusion protein of lung targeted RGD-4C-His Tag-AGAP was expressed successfully.
出处
《微生物学杂志》
CAS
CSCD
2011年第1期15-18,共4页
Journal of Microbiology
基金
国家自然科学基金(30772738)
沈阳市科技局沈阳市药学生物技术重点实验室建设基金(1081102-1-00)
关键词
东亚钳蝎
抗癌镇痛肽
毕赤酵母
真核表达
East Asian pincers scorpion（Buthus martensii Karsch）
AGAP
Pichia pastoris
eukaryotic expression
