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Cloning, Expression, and Purification of the Human Synthetic Survivin Protein in Escherichia coli Using Response Surface Methodology (RSM)

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Abstract

Survivin is one of the novel members of the apoptosis inhibitor protein family in humans. The main activity of the Survivin protein is to suppress caspases activity resulting in negative regulation of apoptosis. Survivin protein can be a potential target for the treatment of cancers between cancerous and normal cells. In the present research, the synthetic Survivin gene with PelB secretion signal peptide was cloned into a prokaryotic expression vector pET21a. The recombinant plasmid pET21a-PelB-Surv was expressed in Escherichia coli (E.coli) BL21, and the relative molecular mass of expressed protein was calculated 34,000 g/mol, approximately. The recombinant protein was purified through chromatography column and characterized by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Response surface methodology (RSM) was used to design 20 experiments for optimization of IPTG concentration, post-induction period, and cell density of induction (OD600). The optimum levels of the selected parameters were successfully determined to be 0.28 mM for IPTG concentration, 10 h for post-induction period, and 3.40768 for cell density (OD600). These findings resulted in 4.14-fold increases in the Survivin production rate of optimum expression conditions (93.6363 mg/ml).

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was funded by the Agricultural and Natural Resources and Education Research Center (AREEO), Kerman, Iran.

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  1. Department of Animal Science Researches, Agriculture and Natural Resources Education and Research Center of Kerman, Agriculture and Natural Resources Education and Research Organization (AREEO), Kerman, Iran

    Nadi Rostami & Laleh Yazdanpanah Goharrizi

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  1. Nadi Rostami
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LYG participated in the conceptualization, study design, planning, methodology, and execution of the study. The data gathered were reviewed and edited by NR. All authors read and approved the final manuscript.

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Correspondence to Laleh Yazdanpanah Goharrizi.

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Rostami, N., Goharrizi, L.Y. Cloning, Expression, and Purification of the Human Synthetic Survivin Protein in Escherichia coli Using Response Surface Methodology (RSM). Mol Biotechnol 65, 326–336 (2023). https://doi.org/10.1007/s12033-021-00399-4

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