中国组织工程研究Issue(18)：2928-2932,5.DOI:10.3969/j.issn.2095-4344.2015.18.025
凋亡素原核表达载体构建及活性测定
Construction of a prokaryotic expression vector for apoptin and activity determination
张艳玲 1徐霞 2江露含 2杜晶春2
作者信息
- 1. 广州医科大学附属第五医院，广东省广州市 510700
- 2. 广州医科大学金域检验学院，广东省广州市 510182
- 折叠
摘要
Abstract
BACKGROUND:Apoptin is a protein which is synthesized in vitro or expressed by genetic engineering, without toxic and transformation activity of normal cel s. Apoptin can specifical y induce the apoptosis of tumor cel s and provide the opportunity of inhibiting the growth of cancer. <br> OBJECTIVE:To construct a prokaryotic expression vector for apoptin, optimize the expression conditions, and detect the activity of the purified protein. <br> METHODS:The apoptin gene that had been constructed was cloned into prokaryotic expression vector pET-28b (+), which was transformed into E.coli host bacteria. Apoptin was induced by isopropyl-beta-D-thiogalactoside, and analyzed by polyacrylamide gel electrophoresis. The inhibition activity of apoptin on tumor cel s was detected. <br> RESULTS AND CONCLUSION:Apoptin gene was successful y cloned into pET-28b (+). Apoptin protein was induced to express in form of inclusion body by isopropyl-beta-D-thiogalactoside (0.5 mmol/L) at 26 ℃. And the expression of apoptin with relative molecular mass of about 15 000 was identified by polyacrylamide gel electrophoresis. The target protein was purified by denaturation-renaturation and affinity chromatography, which has pro-apoptotic effect on lung cancer cel s H460 and H1299. The prokaryotic expression vector pET-28b-apoptin is successful y constructed. The apoptin protein with bioactivity is obtained, which al ows further functional study of apoptin.关键词
实验动物/基因病毒载体及相关因子模型/凋亡素蛋白/原核表达/纯化/活性测定Key words
Apoptosis/Lung Carcinoma分类
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张艳玲,徐霞,江露含,杜晶春..凋亡素原核表达载体构建及活性测定[J].中国组织工程研究,2015,(18):2928-2932,5.
