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Expression and Purification of a Recombinant Enterotoxin Protein Using Different E. coli Host Strains and Expression Vectors

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Abstract

Infection by Enterotoxigenic Escherichia coli is a common cause of diarrhea in animals. The development of vaccines against enterotoxins can effectively control the infection. We have previously constructed a recombinant antigen SLS fused by STa, LTB and STb enterotoxin and it showed a high immunogenicity in mice. Herein, we evaluated the expression of SLS in three different E. coli cells with corresponding plasmids. SLS proteins expressed in E. coli BL21 (DE3) and Rosetta-gami B (DE3) were aggregated as inclusion bodies, and the proteins solubility were not obviously promoted in low temperature combined with adjustment of inducer concentration. In contrast, SLS protein with maltose-binding protein (MBP) yielded from TB1 (DE3) cells were partially soluble. After increasing the IPTG concentration in the medium up to 2 mM and incubating at 37 ℃ for 4 h, the soluble protein yield reached the highest level (4.533 mg/0.2 L culture), which was significantly higher than the expression of SLS protein in Rosetta-gami B (DE3) (P < 0.05). Therefore, the TB1-pMAL expression system can be used for mass extraction and purification of SLS antigen prior to measuring its immunogenicity in pregnant mammals.

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Acknowledgements

This work was supported by the National Key Development Program of China [Grant No. 2017YFD0500601]; National Natural Science Foundation of China [Grant No. 31701719]; the Fundamental Research Funds for the Central Universities [Grant No. DUT20LK29].

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Authors and Affiliations

  1. School of Bioengineering, Dalian University of Technology, Dalian, 116024, China

    Hong Zhao, Yongping Xu, Xiaoyu Li, Gen Li, Haofei Zhao & Lili Wang

  2. Dalian SEM Bio-Engineering Technology Co. Ltd., Dalian, 116620, China

    Yongping Xu

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  1. Hong Zhao
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  4. Gen Li
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Contributions

Hong Zhao, Lili Wang, Yongping Xu and Xiaoyu Li conceptualized and designed the experiments. Hong Zhao, Gen Li and Haofei Zhao performed the experiments and acquired the data. Hong Zhao compiled the results and wrote the manuscript. Hong Zhao and Lili Wang improvised the manuscript.

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Correspondence to Lili Wang.

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The manuscript dose not contain experiments using animals. The manuscript does not contain human studies.

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Zhao, H., Xu, Y., Li, X. et al. Expression and Purification of a Recombinant Enterotoxin Protein Using Different E. coli Host Strains and Expression Vectors. Protein J 40, 245–254 (2021). https://doi.org/10.1007/s10930-021-09973-w

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