SMALL SCALE EXPRESSION, SOLUBILIZATION AND CHARACTERIZATION OF CHIKUNGUNYA VIRUS STRUCTURAL PROTEINS

Authors

  • Namrata Dudha Jaypee Institute of Informtion Technology
  • Jyoti Rana Jaypee Institute of Information Technology
  • Reema Gabrani Jaypee Institute of Information Technology
  • Amita Gupta University of Delhi South Campus
  • Vijay Kumar Chaudhary University of Delhi South Campus
  • Sanjay Gupta Jaypee Institute of Information Technology

Abstract

 

 Objective: The severity and spread of Chikungunya fever in absence of effective antiviral therapy presents a serious public health threat. The present investigation aims to generate soluble and purified viral structural proteins that can be utilized to facilitate generation of reagents for development of both diagnostic and therapeutic measures.

Methods: Bacterial expression system was used for optimization of expression and solubilization of structural proteins of CHIKV (Capsid, 6K and envelope proteins 1-3 [E1, E2 and E3]) as fusions with large (GST) and small (His and Strep) tags on a single platform. Affinity chromatography was used for small scale purification of viral proteins.

Result: The effect of different tags, inducer concentrations, temperatures and duration of induction on solubilization of proteins has been optimized and small scale purification of all the structural proteins has been attempted. Utility of these solubilized proteins has been shown by analyzing the interaction of E2 with all the structural proteins using pull down assay.

Conclusion: Small scale purification of all five structural proteins and ectodomains of envelope proteins E1 and E2 has been standardized. The data and reagents generated can be utilized for large scale purification and studying CHIKV biology.

Keywords: Chikungunya virus, Protein expression, Solubilization, Purification.

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Author Biographies

Namrata Dudha, Jaypee Institute of Informtion Technology

Ph.D Scholar

Center for Emerging Disease, Department of Biotechnology

Jyoti Rana, Jaypee Institute of Information Technology

Ph.D Scholar

Center for Emerging Disease, Department of Biotechnology

Reema Gabrani, Jaypee Institute of Information Technology

Assistant Professor

Center for Emerging Disease, Department of Biotechnology

Amita Gupta, University of Delhi South Campus

Assistant Professor

Department of Bioschemistry

Vijay Kumar Chaudhary, University of Delhi South Campus

Professor and Head

Department of Bioschemistry

Sanjay Gupta, Jaypee Institute of Information Technology

Associate Professor

Center for Emerging Disease, Department of Biotechnology

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Published

01-11-2014

How to Cite

Dudha, N., J. Rana, R. Gabrani, A. Gupta, V. K. Chaudhary, and S. Gupta. “SMALL SCALE EXPRESSION, SOLUBILIZATION AND CHARACTERIZATION OF CHIKUNGUNYA VIRUS STRUCTURAL PROTEINS”. Asian Journal of Pharmaceutical and Clinical Research, vol. 7, no. 5, Nov. 2014, pp. 268-71, https://mail.innovareacademics.in/journals/index.php/ajpcr/article/view/2749.

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