凋亡素融合蛋白质高表达菌株的筛选及其产物活性测定

Screening of High Expression Strain and Activity Assay of Apoptin Fusion Protein

目的筛选凋亡素-HBD融合蛋白质高表达的工程菌株,验证其目标产物具有抑制肿瘤细胞增值的活性。方法在不同大肠杆菌宿主中凋亡素融合蛋白质表达,筛选出高表达工程菌,随后诱导表达和纯化凋亡素融合蛋白质,并通过噻唑蓝法(MTT法)检测目标产物对肿瘤细胞的抑制活性。结果凋亡素-HBD融合蛋白质在SG工程菌的表达量较好。MTT法检测表明,凋亡素融合蛋白质有抑制HeLa细胞增值的活性。结论获得一种凋亡素-HBD融合蛋白质的高表达工程菌株,并证实其产物能够进入HeLa细胞并诱导细胞凋亡。

Objective To screen the high expression strain of apoptin-HBD fusion protein and verify the anti-tumor activity of apoptin-HBD fusion protein.Methods The apoptin-HBD fusion protein was expressed in a variety of E.coli strains.The high expression strain of the target protein was screened.The expressed apoptin-HBD fusion protein was further purified and the inhibitory effect on tumor cells of the fusion protein in vitro was tested by MTT assay.Results The apoptin-HBD fusion protein had a higher expression in E.coli SG strain.MTT assay results indicated that the apoptin-HBD fusion protein could inhibit the proliferation of HeLa cells.Conclusion A high expression engineering strain of the apoptin-HBD fusion protein can be obtained and the apoptin-HBD fusion protein can induce the apoptosis of HeLa cells.