Resumo

AGUERO, J.A et al. MOLECULAR CLONING AND EXPRESSION OF THE GENE THAT CODIFIES FOR THE INFECTIOUS BURSAL DISEASE VIRUS STRUCTURAL PROTEIN, VP3. Rev Salud Anim. [online]. 2007, vol.29, n.1, pp. 39-47. ISSN 0253-570X.

Infectious bursal disease virus (IBDV), a member of the Birnaviridae family, is a double stranded RNA virus that causes considerable economic losses in the poultry industry by inducing bursal destruction, immunosuppression and high mortality in young chickens. Viral capsid is formed by two structural proteins, VP2 and VP3. The vp3 gene (870pb) was amplified by PCR from the vector pcDNA3-Poly containing the gene that encodes for virus polyprotein (pVP2-VP4-VP4). The PCR product was cloned into pQE30 vector. The cloning was screened by PCR using vp3 primers. The gene was expressed as a his-tag fusion protein in M15 E. coli strain. The recombinant protein molecular weight was about 32kDa. The protein was recognized by Western-Blot using an antibody against the virus. Using an ion metal affinity chromatography procedure, a 90% pure recombinant product was obtained. In rabbits, high-titer antibodies against the recombinant VP3, able of recognizing both, the recombinant and native VP3, were obtained. The potential use of these antibodies for VP3 or virus polyprotein expression screening in heterologous systems should be further evaluated.

Palavras-chave : VP3; IBDV; capsid; recombinant protein; IMAC.

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