Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System

YANG Lin; LI Guo-Qing; HUANG Bao-Yang; WANG Quan-Zhong; LONG Qi-Xin; WANG Xun-Zhang

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April 2000

YANG Lin, LI Guo-Qing, HUANG Bao-Yang, WANG Quan-Zhong, LONG Qi-Xin and WANG Xun-Zhang. Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System[J]. Virologica Sinica, 2000, 15(2): 143-148.

Citation: YANG Lin, LI Guo-Qing, HUANG Bao-Yang, WANG Quan-Zhong, LONG Qi-Xin, WANG Xun-Zhang. Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System .VIROLOGICA SINICA, 2000, 15(2) : 143-148.

GST融合蛋白在杆状病毒系统中表达的可溶性研究

1.
中山太学生物防治国家重点实验宣、生物医药中心,广州510275

摘要

将构建好的可表达GST融合蛋白的重组病毒AcMNPVOCC-GST6xHisEtp28感染Sf9细胞,一定时间后取感染了病毒的细胞裂解物上清液进行SDSPAGE分析,结果显示53kDa的融合蛋白(GST6xHisEtp28)呈不溶状态。在原有裂解液的基础上,加固体十二烷基肌氨酸钠至终浓度1.5%,并将TritonX100的比例由1%提高到2%。SDSPAGE结果显示至少有1/3的GST6xHisEtp28处于溶解状态,可溶性GST6xHisEtp28经亲合层析,53kDa的目标蛋白得到纯化
- GST融合蛋白
- , 杆状病毒表达系统
- , 可溶性分析

Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System

1.
State key Lab for Biological Control,Biopharmaceutical Center，Zhangshan University，Guangzhou 510275，China

Abstract

Insect Sf 9 cells were infected with constructed recombinant virus AcMNPV OCC - GST 6xHis Etp 28, which could produce GST fusion protein and the supernatant of 72 h pi cell lysate was examined by SDS PAGE. The results showed that GST 6xHis Etp 28 fusion protein of 53 kDa was expressed in insoluble status. After the sodium salt of the alkylanionic detergent sarkosyl was added to insect cell lysis buffer to a final concentration of 1.5%, and the final concentration of TritonX 100 was increased from 1% to 2%, at least 1/3 of GST 6xHis Etp 28 appeared to be soluble, which was examined by SDS PAGE. Then soluble GST 6xHis Etp 28 was purified by affinity chromatography using glutathione agarose.
- GST fusion pro tein
- , Baculovirus expression system
- , Solubilization analysis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Solubilization Analysis of GST Fusion Protein Expressed in Baculovirus System

1. State key Lab for Biological Control,Biopharmaceutical Center，Zhangshan University，Guangzhou 510275，China

Abstract: Insect Sf 9 cells were infected with constructed recombinant virus AcMNPV OCC - GST 6xHis Etp 28, which could produce GST fusion protein and the supernatant of 72 h pi cell lysate was examined by SDS PAGE. The results showed that GST 6xHis Etp 28 fusion protein of 53 kDa was expressed in insoluble status. After the sodium salt of the alkylanionic detergent sarkosyl was added to insect cell lysis buffer to a final concentration of 1.5%, and the final concentration of TritonX 100 was increased from 1% to 2%, at least 1/3 of GST 6xHis Etp 28 appeared to be soluble, which was examined by SDS PAGE. Then soluble GST 6xHis Etp 28 was purified by affinity chromatography using glutathione agarose.