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Fusion Protein Strategy to Increase Expression and Solubility of Hypervariable Region of VP2 Protein of Infectious Bursal Disease Virus in Escherichia coli

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Abstract

Infectious bursal disease is one of the most important viral diseases in the young chickens. VP2 protein is the major host protective immunogen of the virus. A hypervariable region is present in VP2 protein (hvVP2) that contains immunodominant epitops. The high hydrophobicity of hvVP2 region causes protein aggregation in Escherichia coli (E. coli). The objective of the present study was to improve the expression and the solubility of the hvVP2 protein in E. coli. The effects of fusion partners on the solubility of hvVP2 protein were studied. The protein was expressed in forms of unfused and N-terminally fused to GST and NusA. The results showed that the unfused hvVP2 protein was expressed in very low level. But, N-terminally fused hvVP2 protein to GST (glutathione-S-transferase) and NusA (N utilization substance A) showed significantly enhanced protein expression. The fusion of GST and hvVP2 was produced in aggregated form while in the presence of NusA, the hvVP2 protein was expressed in a soluble form. The NusA-hvVP2 protein was detected by a neutralizing monoclonal antibody, 1A6, in antigen-capture ELISA. In conclusion, the NusA protein is a suitable fusion partner to improve expression and solubility of the hvVP2 protein in E. coli.

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Abbreviations

ABTS:

2-2′-azino-di-(3-ethylbenzthiazoline sulfonic acid)

DAB:

3,3′-diaminobenzidine

GST:

Glutathione-S-transferase

HRP:

Horseradish peroxidase

hvVP2:

Hypervariable region of VP2 protein

IBD:

Infectious bursal disease

IBDV:

Infectious bursal disease virus

IMAC:

Immobilized metal-affinity chromatography

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

Ni-NTA:

Ni2+-nitrilotriacetate

NusA:

N utilization substance A

RT-PCR:

Reverse transcription polymerase chain reaction

SD:

Standard deviations

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

vvIBDV:

Very virulent IBDV

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Acknowledgments

The authors thank from National Institute of Genetic Engineering and Biotechnology and Isfahan University of Technology for financial supports.

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Authors and Affiliations

  1. National Institute of Genetic Engineering & Biotechnology (NIGEB), P.O. Box 14155-6343, Tehran, Iran

    Sahar Sadat Sedighzadeh & Mehdi Shamsara

  2. Agricultural Biotechnology Department, Isfahan University of Technology, Isfahan, Iran

    Sahar Sadat Sedighzadeh & Azar Shahpiri

Authors
  1. Sahar Sadat Sedighzadeh
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  2. Mehdi Shamsara
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  3. Azar Shahpiri
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Correspondence to Mehdi Shamsara.

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Sedighzadeh, S.S., Shamsara, M. & Shahpiri, A. Fusion Protein Strategy to Increase Expression and Solubility of Hypervariable Region of VP2 Protein of Infectious Bursal Disease Virus in Escherichia coli . Protein J 31, 580–584 (2012). https://doi.org/10.1007/s10930-012-9437-2

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  • DOI: https://doi.org/10.1007/s10930-012-9437-2

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