| 重组人可溶性TRAIL在毕赤酵母中表达与纯化及其抗肿瘤细胞活性 |
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| 引用本文： | 黄晓平,王晓,董浩,赵小峰,李招发,王启钊,许瑞安,刁勇.重组人可溶性TRAIL在毕赤酵母中表达与纯化及其抗肿瘤细胞活性[J].中国生化药物杂志,2010,31(2). |
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| 作者姓名： | 黄晓平 王晓 董浩 赵小峰 李招发 王启钊 许瑞安 刁勇 |
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| 作者单位： | 1. 华侨大学,分子药物学研究所,福建,泉州,362021
2. 华侨大学,分子药物学研究所,福建,泉州,362021;华侨大学,分子药物教育部工程研究中心,福建,泉州,362021 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，国家科技重大专项课题 |
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| 摘 要： | 目的研究重组人可溶性TRAIL(sTRAIL)蛋白的表达和纯化,以及对HepG2细胞增殖与凋亡的影响。方法将人sTRAIL基因插入毕赤酵母表达载体pPIC9中,构建甲醇酵母分泌型表达载体pPIC9-sTRAIL,电击转化至GS115(his4),MD平板筛选出阳性菌株。对工程菌株发酵培养基成分、pH、甲醇浓度、诱导表达时间等条件进行优化,并初步分析重组sTRAIL蛋白的体外抗肿瘤活性。结果工程菌株在BMMY培养基(pH6.0)、1%甲醇条件下诱导表达48h目的蛋白浓度最高可达(58.7±2.4)mg/L。重组人sTRAIL蛋白能抑制HepG2细胞的增殖、诱导HepG2细胞的凋亡。结论优化了人sTRAIL的表达和纯化工艺条件,所生产的sTRAIL可用于抗肝癌新药的开发。
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| 关 键 词： | sTRAIL 毕赤酵母 表达 纯化 肿瘤 |
Expression and purification of human soluble TRAIL in Pichia pastoris and its anti-tumor activity in vitro |
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| HUANG Xiao-ping,WANG Xiao,DONG Hao,ZHAO Xiao-feng,LI Zhao-fa,WANG Qi-zhao,XU Rui-an,DIAO Yong.Expression and purification of human soluble TRAIL in Pichia pastoris and its anti-tumor activity in vitro[J].Chinese Journal of Biochemical Pharmaceutics,2010,31(2). |
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| Authors: | HUANG Xiao-ping WANG Xiao DONG Hao ZHAO Xiao-feng LI Zhao-fa WANG Qi-zhao XU Rui-an DIAO Yong |
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| Institution: | HUANG Xiao-ping1,WANG Xiao1,DONG Hao1,ZHAO Xiao-feng1,LI Zhao-fa1,2,WANG Qi-zhao1,XU Rui-an1,DIAO Yong1,2(1.Institute of Molecular Medicine,Huaqiao University,2.Molecular Medicine Engineering ResearchCenter of the Ministry of Education,Quanzhou 362021,China) |
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| Abstract: | Purpose To investigate human soluble TRAIL (sTRAIL) protein expression and purification and its potential anti-tumor activity on hepatocellular carcinoma (HepG2).Methods Soluble TRAIL gene ligated with expression vector pPIC9 was transfected into GS115 (his4) and the recombination strain expressing sTRAIL was screened by MD plate.The effects of different media,methanol inducement period,methanol concentration,and pH were investigated and optimized using shaking flask.The anti-tumor activity of sTRAIL with H... |
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| Keywords: | sTRAIL sTRAIL Pichia pastoris expression purification tumor |
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