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Vaccination Against Hydatidosis: Molecular Cloning and Optimal Expression of the EG95NC− Recombinant Antigen in Escherichia coli

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Abstract

Cystic echinococcosis (CE) is a widely distributed zoonosis that is highly endemic in the Mediterranean basin. The disease represents a serious public health threat and causes economic losses. The parasite life-cycle involves dogs and ruminants as definitive and intermediate hosts; humans are accidently infected, causing serious clinical issues. Vaccination of ruminants and dog treatments represent the most efficient measures to prevent parasite transmission. The recombinant protein vaccine, EG95, has been used successfully in sheep vaccine trials against CE in several countries. In this study, we expressed the modified antigen, EG95NC-GST, in Escherichia coli for use as a vaccine against Echinococcus granulosus in ruminants. We tested three different media formulations for E. coli culture and established for each culture conditions for optimal levels of soluble EG95 expression. The results demonstrate that SOC and TB media provided high yields in cell density and EG95 protein expression. Purification of the recombinant protein with affinity chromatography (using FPLC) was also performed to increase the purity of the EG95NC−-GST antigen.

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Abbreviations

CE:

Cystic echinococcosis

GST:

Glutathione S-transferase

IPTG:

Isopropyl-β-d-thiogalactoside

LB:

Luria broth

TB:

Terrific broth

SOC:

Super broth

FPLC:

Fast protein liquid chromatography

SDS-PAGE:

Sodium dodecyl sulphate-polyacrylamide gel Electrophoresis

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Funding

This work was funded by MCI animal health company.

Author information

Author notes

  1. M. M. Ennaji and M. Elharrak have contributed equally.

Authors and Affiliations

  1. Research and Development of Recombinant Vaccine, Multi-Chemical Industry, Lot. 157, Z I, Sud-Ouest (ERAC) B.P.: 278, 28810, Mohammedia, Morocco

    M. Jazouli, K. Tadlaoui, A. Belmlih & M. Elharrak

  2. Molecular Parasitology Laboratory, The University of Melbourne, Princes Highway, Werribee, VIC, 3030, Australia

    M. Jazouli, M. Lightowlers & C. G. Gauci

  3. Laboratory of Virology, Hygiene and Microbiology, Faculty of Science and Technology, University Hassan II-Casablanca, 20650, Mohammedia, Morocco

    M. Jazouli & M. M. Ennaji

Authors
  1. M. Jazouli
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  2. M. Lightowlers
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  3. C. G. Gauci
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  4. K. Tadlaoui
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  5. A. Belmlih
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  6. M. M. Ennaji
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  7. M. Elharrak
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Contributions

MH, KT and ML initiated the study, participated in experimental design. MJ conducted the experiment. All authors participated in data analysis; interpretation of the results, MJ, MH wrote the paper and ML review it. All authors read and approved the final manuscript.

Corresponding author

Correspondence to M. Jazouli.

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Conflict of interest

The authors declare that they have no competing interests.

Ethical Approval

No human or animal subjects were used in this study. Declarations of ethical treatment of human subjects or animals, or consent of human subjects are therefore not applicable.

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Jazouli, M., Lightowlers, M., Gauci, C.G. et al. Vaccination Against Hydatidosis: Molecular Cloning and Optimal Expression of the EG95NC− Recombinant Antigen in Escherichia coli . Protein J 36, 472–477 (2017). https://doi.org/10.1007/s10930-017-9742-x

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  • DOI: https://doi.org/10.1007/s10930-017-9742-x

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