Abstract
37-kDa immature laminin receptor protein (iLRP), the precursor of 67-kDa laminin receptor protein (LRP), is overexpressed on the surface of most cancer cells and recognized as a universal tumor antigen. The role makes it a potential target for cancer immunotherapy, which has been well-studied. Our study aimed to produce high quality of human iLRP in bacteria so that the needs in research of its clinical application could be met. The powerful system for heterologous protein expression, pET system was used. Two types of DNA sequences encoding the same amino acid sequences were separately cloned into the vector pET30a(+). One of the resulting vectors includes the wild-type iLRP, and other one includes the codon-optimized iLRP. The expression by both genes was then compared in Escherichia coli BL21(DE3). Our results revealed that the performance of codon optimization was crucial for the expression of human iLRP in Escherichia coli. The yield was significantly enhanced up to 300 mg/L of bacterial culture by this approach.
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Acknowledgements
This study was supported by the National Natural Science Foundation of China (Grant no. 81560276), and the Joint Funds of Science and Technology Department of Guizhou Province of China (Grant No. LKZ[2013]02), and the Foundation for Returnees from the Department of Human Resources and Social Security of Guizhou Province of China (Grant no. [2014]08).
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Dr. BL conceived and designed the study and performed the data analysis and wrote the manuscript; QK designed and performed the experiments and data analysis; DZ, and LY helped perform the analysis with constructive discussions.
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On behalf of all authors, Dr. Bainan Liu states that there is no conflict of interest.
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Liu, B., Kong, Q., Zhang, D. et al. Codon optimization significantly enhanced the expression of human 37-kDa iLRP in Escherichia coli. 3 Biotech 8, 210 (2018). https://doi.org/10.1007/s13205-018-1234-y
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DOI: https://doi.org/10.1007/s13205-018-1234-y