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| CLONING SEGMENT SPIKE PROTEIN GENE OF SARS-COV AND ITSEXPRESSION IN ESCHERICHIA COLI | |
| 作者姓名： | 刘中华 许文波 毛乃颖 张燕 朱贞 崔爱利 杨建国 胡海涛 |
| 作者单位： | [1]DepartmentofAnatomy,MedicalSchoolofXi＇anJiaotongUniversity,Xi＇an710061,China [2]NationalMeaslesLab,NationalInstituteforViralDiseaseControlandPrevention,ChinaCDC,Beijing100050,China |
| 摘 要： | Objective Expressing and purifying the seglnent of SARS-CoV spike protein in E. Coli Methods The target gene was obtained by RT-PCR. The PCR product was cloned into pEGM- T Easy Vector, sequencing and double restriction digestion (BamH Ⅰ , Pst Ⅰ) were performed. The target gene was subcloned into PQE30 expression vector. The gene was expressed in the E. coli strain M15 cells induced by IPTG. The protein was purified with a nickel HiTrap chelating metal affinity column. Results The recombinant expression plasmid was successfully constructed and the protein was well expressed in E. coli strain M15 cells. The ideal pure protein was obtained by purification. Western blotting analysis suggested the protein could act with the convalescent sera of lab confirmed SARS patients. Conclusion The segment of SARS-CoV spike protein was well expressed and purified, and can be applied in diagnosis and immunological research of SARS. |
| 关 键 词： | 非典型性肺炎 穗状蛋白质 蛋白质提纯 蛋白质表述 |
CLONING SEGMENT SPIKE PROTEIN GENE OF SARS-COV AND ITS EXPRESSION IN ESCHERICHIA COLI |
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| Liu Zhonghua #,Xu Wenbo,Mao Naiying,Zhang Yan,Zhu Zhen,Cui Aili,Yang Jianguo,Hu Haitao# #.CLONING SEGMENT SPIKE PROTEIN GENE OF SARS-COV AND ITSEXPRESSION IN ESCHERICHIA COLI[J].Academic Journal of Xi’an Jiaotong University,2004,16(1):50-53. | |
| Authors: | Liu Zhonghua # Xu Wenbo Mao Naiying Zhang Yan Zhu Zhen Cui Aili Yang Jianguo Hu Haitao# # |
| Institution: | 1. National Measles Lab, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100050, China;Department of Anatomy, Medical School of Xi'an Jiaotong University, Xi'an 710061,China 2. National Measles Lab, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100050, China 3. Department of Anatomy, Medical School of Xi'an Jiaotong University, Xi'an 710061,China |
| Abstract: | Objective Expressing and purifying the segment of SARS-CoV spike protein in E.Coli. Methods The target gene was obtained by RT-PCR. The PCR product was cloned into pEGM- T Easy Vector, sequencing and double restriction digestion ( BamHⅠ,PstⅠ) were performed. The target gene was subcloned into PQE30 expression vector. The gene was expressed in the E.coli strain M15 cells induced by IPTG. The protein was purified with a nickel HiTrap chelating metal affinity column. Results The recombinant expression plasmid was successfully constructed and the protein was well expressed in E. coli strain M15 cells. The ideal pure protein was obtained by purification. Western blotting analysis suggested the protein could act with the convalescent sera of lab confirmed SARS patients. Conclusion The segment of SARS-CoV spike protein was well expressed and purified, and can be applied in diagnosis and immunological research of SARS. |
| Keywords: | SARS-CoV spike protein expression purification |
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