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- 2017
R9-FOXM1(1-234aa)重组蛋白批量纯化及其抑瘤效应研究Keywords: 多聚精氨酸 细胞穿膜肽 FOXM1 肿瘤治疗arginine-rich cell-penetrating peptides FOXM1 tumor therapy Abstract: 通过基因工程构建重组表达质粒pET15b-R9-FOXM1(1-234aa),转化大肠杆菌建立构建表达R9-FOXM1(1-234aa)的菌株.采用原核表达系统和His-tag亲和纯化手段,规模化制备纯化穿膜肽R9-FOXM1(1-234aa),获得的蛋白的纯度达到90%以上.用R9-FOXM1(1-234aa)穿膜肽处理不同的肿瘤细胞,通过MTT实验研究其细胞效应.结果显示:当R9-FOXM1(1-234aa)穿膜肽的浓度达到2 mM时,肿瘤细胞的死亡率为50%左右.实验表明穿膜肽R9-FOXM1(1-234aa)抑制不同肿瘤细胞的生长,有可能成为治疗肿瘤的潜在蛋白类药物.A recombinant protein expression vector pET15b-R9-FOXM1(1-234aa) was constructed and transformed to E. coli in order to generate a strain expressing R9-FOXM1(1-234aa). The recombinant protein R9-FOXM1 (1-234aa) (R9-FOXM1(1-234aa)) was isolated at a large scale through His-tag affinity chromatography. The purity of the purified protein reached 90%. Moreover, MTT assay was used to test the effect of R9-FOXM1(1-234aa) on cells, and the test results showed that R9-FOXM1(1-234aa) caused the cell death of different types of cancer cells with a half lethal dose around 2 mm。 The results also demonstrated that R9-FOXM1(1-234aa) suppressed the proliferation of cancer cells and may be considered as a potential angent for anti-cancer in the future.
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