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| Expression and Purification of GST-GFMCry11B Fusion Protein in E.coli and Preparation of Polyclonal Antibody Against GFMCry11B |
| DING Min1,2 WANG Feng1** SU Jun1 CHEN Zhi-Hou1 CHEN Zai-Jie1 SUN Ming2 |
(1. Fujian Province Key Laboratory of Genetic Engineering for Agriculture, Fujian Academy of Agricultural Sciences.
Fuzhou 350003, China; 2. Key Laboratory of Agriculture Microbiology, Ministry of
Agriculture, Huazhong Agricultural University, Wuhan 430070, China) |
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Abstract Abstract: The coding region of GFM cry11B, fully synthesized gene, was cloned into the multi-clone sites of pGEX4T vector to produce the recombined expression plasmid pGC by the methods of restriction enzyme digestion. The recombinant plasmid was transferred into Escherichia coli BL21(DE3). GST-GFMCry11B fusion protein was obtained after adding IPTG into the growth media. SDS-PAGE analysis revealed that the fusion protein was highly expressed and accumulated up to above 21% of the total bacterial protein and the main expression production was deposited as inclusion body. The GST-GFMCry11B was purified with Glutathione Sephrose 4B after soluble treatment. GFMCry11B protein cleaved by Thrombin was collected and used as the antigen to immune the rabbits. ELISA assay showed that the titer of the prepared polyclonal antibody was 1∶8000 and had high specialties.
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Received: 01 January 1900
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Corresponding Authors:
WANG Feng
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