Cloning and Codon-optimized Expression of Structural Protein Hypervariable Region of VP2 from Infectious Bursal Disease Virus

International Journal of Molecular and Clinical Microbiology

مقاله 2، دوره 2، شماره 1، مهر 2012، صفحه 119-123   

نوع مقاله: Research Article

نویسندگان

Sahar sadat Sedighzadeh¹؛ Mehdi Shamsara^{* 2}؛ Soheil Haji Khodadad³؛ Ayatollah Nasrollahi Omran⁴

¹Animal, Avian and Marine Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran. Agricultural Biotechnology Department, Isfahan University of Technology, Isfahan, Iran.

²Animal, Avian and Marine Biotechnology Department, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

³Microbiology Department, Islamic Azad University - Tonekabon Branch, Tonekabon, Iran.

⁴Medical Mycology Department, Faculty of Medical Sciences, Azad University-Tonekabon Branch, Tonekabon, Iran

چکیده

Infectious bursal disease virus (IBDV) is the causative agent of Gumboro disease, an infectious disease of global economic importance in poultry. Structural protein VP2 of IBDV is the most frequently studied protein due to its significant roles in virus attachment, protective immunity, and serotype specificity. The objective of the present study was to improve the expression of hypervariable region of VP2 protein (hvVP2) in Escherichia  coli (E.coli). The results showed that the hvVP2 was expressed in very low amount in E.coli. But, codon optimized hvVP2 protein showed significantly enhanced protein expression level. The coding sequence of hvVP2 was amplified and then identified by polymerase chain reaction (PCR) and sequencing. To achieve high-level expression of hvVP2 protein, we optimized hvVP2 gene base on E. coli preferred codons and synthesized the optimized gene. The synthetical gene was cloned into expression vector pET-26b       and expressed in E.coli BL21 (DE3). After induction with Isopropyl-D-1-Thiogalactopyranoside (IPTG) and optimization the conditions of expression, the hvVP2 protein was relatively increased and identified by SDS-PAGE and Western blotting. Productive conformation can now be used for structure-based design purposes as well as structure-function relation of VP1 protein. It is suggested that the codon optimized hvVP2-His protein may be a useful option (but it is not enough) for developing diagnostic tests and immunization proposes.

کلیدواژه‌ها

hvVP2؛ IBDV؛ codon-optimized؛ protein expression