3 Biotech

, 8:210 | Cite as

Codon optimization significantly enhanced the expression of human 37-kDa iLRP in Escherichia coli

Short Reports

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.

Keywords

Codon optimization Immature laminin receptor protein Heterologous expression pET system 

Notes

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).

Author contributions

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.

Compliance with ethical standard

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of interest

On behalf of all authors, Dr. Bainan Liu states that there is no conflict of interest.

Supplementary material

13205_2018_1234_MOESM1_ESM.pdf (1.3 mb)
Supplementary material 1 (PDF 1318 kb)

References

  1. Barsoum AL, Schwarzenberger PO (2014) Oncofetal antigen/immature laminin receptor protein in pregnancy and cancer. Cell Mol Biol Lett 19:393–406.  https://doi.org/10.2478/s11658-014-0203-7 CrossRefGoogle Scholar
  2. Barsoum AL, Liu B, Rohrer JW, Coggin JH Jr, Tucker JA, Pannell LK, Schwarzenberger PO (2009) Production, safety and antitumor efficacy of recombinant oncofetal antigen/immature laminin receptor protein. Biomaterials 30:3091–3099.  https://doi.org/10.1016/j.biomaterials.2009.02.022 CrossRefGoogle Scholar
  3. Burgess-Brown NA, Sharma S, Sobott F, Loenarz C, Oppermann U, Gileadi O (2008) Codon optimization can improve expression of human genes in Escherichia coli: a multi-gene study. Protein Expr Purif 59:94–102.  https://doi.org/10.1016/j.pep.2008.01.008 CrossRefGoogle Scholar
  4. Chung BK, Lee DY (2012) Computational codon optimization of synthetic gene for protein expression. BMC Syst Biol 6:134.  https://doi.org/10.1186/1752-0509-6-134 CrossRefGoogle Scholar
  5. Coggin JH Jr, Barsoum AL, Rohrer JW (1999) 37 kiloDalton oncofetal antigen protein and immature laminin receptor protein are identical, universal T-cell inducing immunogens on primary rodent and human cancers. Anticancer Res 19:5535–5542Google Scholar
  6. DiGiacomo V, Meruelo D (2016) Looking into laminin receptor: critical discussion regarding the non-integrin 37/67-kDa laminin receptor/RPSA protein. Biol Rev Camb Philos Soc 91:288–310.  https://doi.org/10.1111/brv.12170 CrossRefGoogle Scholar
  7. Friedrichs B et al (2008) Humoral immune responses against the immature laminin receptor protein show prognostic significance in patients with chronic lymphocytic leukemia. J Immunol 180:6374–6384CrossRefGoogle Scholar
  8. Gao CY, Xu TT, Zhao QJ, Li CL (2015) Codon optimization enhances the expression of porcine beta-defensin-2 in Escherichia coli. Genet Mol Res 14:4978–4988.  https://doi.org/10.4238/2015.May.12.1 CrossRefGoogle Scholar
  9. Gvritishvili AG, Leung KW, Tombran-Tink J (2010) Codon preference optimization increases heterologous PEDF expression. PLoS ONE 5:e15056.  https://doi.org/10.1371/journal.pone.0015056 CrossRefGoogle Scholar
  10. Hanson G, Coller J (2018) Codon optimality, bias and usage in translation and mRNA decay. Nat Rev Mol Cell Biol 19:20–30.  https://doi.org/10.1038/nrm.2017.91 CrossRefGoogle Scholar
  11. McClintock SD et al (2015) Monoclonal antibodies specific for oncofetal antigen–immature laminin receptor protein: effects on tumor growth and spread in two murine models. Cancer Biol Ther 16:724–732.  https://doi.org/10.1080/15384047.2015.1026484 CrossRefGoogle Scholar
  12. Mierendorf RC, Morris BB, Hammer B, Novy RE (1998) Expression and purification of recombinant proteins using the pET system. Methods Mol Med 13:257–292.  https://doi.org/10.1385/0-89603-485-2:257 Google Scholar
  13. Nguyen AN et al (2017) Prokaryotic soluble expression and purification of bioactive human fibroblast growth factor 21 using maltose-binding protein. Sci Rep 7:16139.  https://doi.org/10.1038/s41598-017-16167-x CrossRefGoogle Scholar
  14. Poon SL, Klausen C, Hammond GL, Leung PC (2011) 37-kDa laminin receptor precursor mediates GnRH-II-induced MMP-2 expression and invasiveness in ovarian cancer cells. Mol Endocrinol 25:327–338.  https://doi.org/10.1210/me.2010-0334 CrossRefGoogle Scholar
  15. Rohrer JW, Barsoum AL, Coggin JH Jr (2006) Identification of oncofetal antigen/immature laminin receptor protein epitopes that activate BALB/c mouse OFA/iLRP-specific effector and regulatory T cell clones. J Immunol 176:2844–2856CrossRefGoogle Scholar
  16. Shi Y, Halperin SA, Lee SF (2018) Expression, purification, and functional analysis of an antigen-targeting fusion protein composed of CD40 ligand and the C-terminal fragment of ovalbumin. Protein Expr Purif 142:37–44.  https://doi.org/10.1016/j.pep.2017.09.015 CrossRefGoogle Scholar
  17. Shin CS, Hong MS, Bae CS, Lee J (1997) Enhanced production of human mini-proinsulin in fed-batch cultures at high cell density of Escherichia coli BL21(DE3)[pET-3aT2M2]. Biotechnol Prog 13:249–257.  https://doi.org/10.1021/bp970018m CrossRefGoogle Scholar
  18. Song H, Li G, Mai W, Huang G, Chen K, Zhou Y, Chen H (2014) Codon optimization enhances protein expression of Bombyx mori nucleopolyhedrovirus DNA polymerase in E. coli. Curr Microbiol 68:293–300.  https://doi.org/10.1007/s00284-013-0476-5 CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of ImmunologyZunyi Medical UniversityZunyiChina

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