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Cloning, expression, and purification of fusion proteins encoded by
19kD-ESAT6 in Mycobacterium tuberculosis |
| SHI Li1； DING Yuan-Sheng2；YANG Hua2； LIU Yao-Ting1；LIU Zhong-Hua2；HU Zhong-Yi2；HUANG Rui1 |
| 1. Department of Microbiology, Medical College of Soochow University, Suzhou 215123, China; 2. Shanghai Tuberculosis Key Laborarory, The Pulmonary Hospital Affiliated to Tongji University, Shanghai 200433, China |
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Abstract The genes encoding 19kD lipoprotein and early secreting antigenic target-6( ESAT-6) were amplified from genome of the standard Mycobacterium tuberculosis H37Rv using polymerase chain reaction, and then cloned into the vector pMD18-T followed by subcloning into the expression vector pET-21a, respectively. Recombinant 19kD-ESAT6 was expressed in E. coli, and then purified by Ni-NTA. The antigenicity of the fusion protein was analyzed by Western blot analysis. The recombinant 19kD-ESAT6 plasmid was constructed successfully, and could be expressed efficiently in E. coli BL21 ( DE3) . The relative molecular mass of the fusion protein was approximately 29 ×103 by SDS-PAGE. The recombinant 19kD-ESAT6 protein showed specific antigenicity as determined by Western blot, and can be used for the development of serodiagnostic reagents.
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Received: 01 January 1900
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Corresponding Authors:
HU Zhong-Yi；HUANG Rui
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2009, Vol. 4 
