| Expression and Purification of SARS Coronavirus Membrane Protein |
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| 作者姓名： | 戴五星 雷明军 吴少庭 陈智浩 梁靓 潘晖榕 秦莉 高士同 袁仕善 张仁利 |
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| 作者单位： | [1]DepartmentofBiochemistryandMolecularBiology,SchoolofBasicMedicalScience,TongjiMedicalCollege,HuangzhongUniversityofScienceandTechnology,Wuhan430040,China [2]ShenzhenMunicipalCenterforDiseaseControlandPrevention,Shenzhen518020,China |
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| 摘 要： | To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants were transformed into Escherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-β-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0. 992 1 ku) protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further study on SARS virus vaccine and diagnostic agents.
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| 关 键 词： | SARS DNA Western-blot RT-PCR SDS-PAGE BL IPTG |
| 收稿时间： | 26 December 2003 |
Expression and purification of SARS coronavirus membrane protein |
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| Dai?Wuxing,Lei?Mingjun,Wu?Shaoting,Chen?Zhihao,Liang?Liang,Pan?Hujrong,Qin?Li,Gao?Shitong,Yuan?Shishan,Zhang?Renli.Expression and Purification of SARS Coronavirus Membrane Protein[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2004,24(5):414-416. |
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| Authors: | Dai Wuxing Lei Mingjun Wu Shaoting Chen Zhihao Liang Liang Pan Hujrong Qin Li Gao Shitong Yuan Shishan Zhang Renli |
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| Institution: | 1. Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Tongji Medical College, Huangzhong University of Science and Technology, Wuhan 430040, China
2. Shenzhen Municipal Center for Disease Control and Prevention, Shenzhen 518020,China |
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| Abstract: | Summary To construct a recombinant plasmid Pet23a-M, the gene encoding severe acute respiratory syndrome (SARS) coronavirus membrane
protein was amplified by RT-PCR and cloned into the expression plasmid Pet23a. Results of restriction endonuclease analysis,
PCR detection and DNA sequencing analysis revealed that the cloned DNA sequence was the same as that reported. The recombinants
were transformed intoEscherichia coli (E. Coli) BL21 (DE3) and induced by Isopropyl-β-D-thiogalactopyranoside (IPTG). The expression of 27 kD (1 kD=0.992 1 ku)
protein was detected by SDS-PAGE and pured by metal chelated chromatography. Results of Western-blot showed that this expressed
protein could react with antibodies in sera of SARS patients during convalescence. This provided the basis for the further
study on SARS virus vaccine and diagnostic agents.
DAI Wuxing, male, born in 1950. Associate Professor |
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| Keywords: | SARS membrane protein gene expression protein purification Western-blot |
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