文档介绍：1
食品与药品 Food and Drug 2012年第14卷第01期 5
凋亡素融合蛋白质高表达菌株的筛选及其产物活性测定
王焕焕1,汪雅雯1,曹晨1,赵健1,陆玉婷2,王富军2*
(1. 华东理工大学生物反应器工程国家重点实验室,上海 200237;2. 浙江日升昌药业有限公司,浙江东阳
市 322100)
摘要: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。
关键词:凋亡素-HBD;高表达工程菌;噻唑蓝法
中图分类号: 文献标识码:A 文章编号:1672-979X(2012)01-0005-03
Screening of High Expression Strain and Activity Assay of Apoptin Fusion Protein
WANG Huan-huan1, WANG Ya-wen1, CAO Chen1, ZHAO Jian1, LU Yu-ting2, WANG Fu-jun2
(1. State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai
200237, China; 2. Zhejiang Reachall Pharmaceutical Co., Ltd, Dongyang 322100, China)
Abstract: 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. co