Revista MVZ Córdoba
versão impressa ISSN 0122-0268
Objective. To construct a recombinant plasmid expressing the ASP1r protein of A. caninum and evaluate its immunogenic capacity in a murine model. Materials and methods. RNA of adults of Ancylostoma caninum was amplified by reverse transcription polymerase chain reaction. The ASP1 protein gene was inserted into the pcDNA3 vector. Plasmid was digested with Bamh1 and EcoR1 and cloning was performed directionally. Later a transformation and selection of E. coli DH5a cell competent with the product for the ligation was made. Then, a screening by PCR was carried out to confirm the presence of ASP1 gene. PcDNA3-ASP1 was inoculated by intraglandular parotide and intramuscular route in Balb/c mice. Antibodies in these animals was measured in serum and saliva by ELISA and immunochemistry. Results. PcDNA3-ASP1 was incorporated and expressed in E. coli DH5a cell. This recombinant plásmid was able to produce antibodies anti-ASP1 specific in Balb/c mice. Discussion. It was possible to demonstrate that using of pcDNA3-ASP1 did not display reactogenicity and it did not produce unfavorable reactions, futhermore, it induced a humoral response against the excreción/secretion protein of Ancylostoma caninum in mice.
Palavras-chave : Ancylostoma caninum; ASP1 protein; immunohistochemistry; DNA vaccines; Cloning Molecular.