Resumo

AGUERO, J.A et al. In vitro EXPRESSION IN Escherichia coli OF THE vlhA.5.02 GENE FRAGMENT, A MEMBER OF THE MAJOR HEMAGLUTININ FAMILY OF Mycoplasma gallisepticum. Rev Salud Anim. [online]. 2011, vol.33, n.1, pp. 32-37. ISSN 0253-570X.

Mycoplasma gallisepticum, the causal agent of the chronic respiratory disease of chickens, is the most economically significant for poultry industry. An 800pb DNA fragment of Mycoplasma gallisepticum which was amplified from the recombinant vector pTTQ18 was cloned into the expression vector pQE-32. The polypeptide was expressed as a fusion protein to 6X-histidine tag; hence its final size was 31kDa. It was detected by Western blott being recognized by a polyclonal antiserum to M. gallisepticum, as well as by a monoclonal antibody to histidine. The polypeptide was purified by nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity cromatography yielding a final product of nearly 90% of purity.

Palavras-chave : Mycoplasma gallisepticum; cloning; expression; purification; PCR; nickel-nitrilotriacetic acid (Ni-NTA) metal-affinity chromatography.

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