Expression in E ．Coti and Antigenicity of HGV NS5 Protein

GONG Rui; MA Hul—wen; TONG Li—heng

PDF
Cite
Share

facebook

twitter

google

linkedin

April 2001

GONG Rui, MA Hul—wen and TONG Li—heng. Expression in E ．Coti and Antigenicity of HGV NS5 Protein[J]. Virologica Sinica, 2001, 16(2): 114-118.

Citation: GONG Rui, MA Hul—wen, TONG Li—heng. Expression in E ．Coti and Antigenicity of HGV NS5 Protein .VIROLOGICA SINICA, 2001, 16(2) : 114-118.

庚型肝炎病毒NS5 cDNA片段的表达及其免疫原性的研究

1.
1武设大学生命科学学院，湖北武汉430072：2 武汉市生物技术研究开发中心．湖北武汉430033

摘要

一段长度为880 bp的庚型肝炎病毒eDNA在太肠杆菌BL21(DE3)菌株中得到表达。此eDNA被插入到表达质粒pGEX-5X-I中，位于编码日本血吸虫答胱甘肽硫转移酶(GST)的DNA序列下游，并与GST处于同一阅读框。用乳糖在37’C下诱导表达出以包涵体形式存在的GST-NS53融合蛋白，并用脲溶法提取了该蛋白；在20℃诱导时，表达出的蛋白大部分可溶．用各胱甘肽Sepharose 4B亲和层析柱对可溶性的融合蛋白进行了纯化。免疫印迹实验证明，此融合蛋白能被庚型肝炎病人的血清和自制的抗GST血清特异性地识别。用PC gene软件对NS53氨基酸序列的亲水性和抗原决定簇进行了分析。本研究为庚型肝炎ELISA诊断试剂研制打下了基础。
- 庚肝病毒非结构蛋白i基因表达
- , 免疫原性

Expression in E ．Coti and Antigenicity of HGV NS5 Protein

1.
1．College of Life Sciences，Wuhan University，Wuhan 430072，China2 Wuhan R＆D Center for Biotechnology，Wuhan 430033，China

Abstract

880 bp cD A localized to the putative NS5 region of HGV genome was expressed in E． colf BL2I(DE3) The eDNA fragment was inserted into a plasmid pGEX-5X一1，at the downstream of the DNA sequence encoding Schistosoma japonicum glutathioue S_transferase(GST)，in the S~TIe reading frame with the gene of GST．A 60KD GS丁_ S53 fusion protein was expressed at 37℃ in a forill of nclusion bodies amounting to 30 percent of total host protein whereas at 20℃ mainly in a form of soluble pmtein The fusion protein was extracted and purified to homologue，The purified GST— NS53 fusion protein could be specifically recognized with either the sera from the patient infected by HGV or the antisera directed against GST
- HGV NS5 pm tein
- , Gene expression

References
Proportional views

PDF

Article Metrics

Article views(3891) PDF downloads(905) Cited by(0)

Proportional views

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

Expression in E ．Coti and Antigenicity of HGV NS5 Protein

1. 1．College of Life Sciences，Wuhan University，Wuhan 430072，China2 Wuhan R＆D Center for Biotechnology，Wuhan 430033，China

Abstract: 880 bp cD A localized to the putative NS5 region of HGV genome was expressed in E． colf BL2I(DE3) The eDNA fragment was inserted into a plasmid pGEX-5X一1，at the downstream of the DNA sequence encoding Schistosoma japonicum glutathioue S_transferase(GST)，in the S~TIe reading frame with the gene of GST．A 60KD GS丁_ S53 fusion protein was expressed at 37℃ in a forill of nclusion bodies amounting to 30 percent of total host protein whereas at 20℃ mainly in a form of soluble pmtein The fusion protein was extracted and purified to homologue，The purified GST— NS53 fusion protein could be specifically recognized with either the sera from the patient infected by HGV or the antisera directed against GST