DNA vaccine using syntetic gene derivate of peptide SBm 7462 anti-tick Rhipicephalus (Boophilus) microplus and assessment of immune response in Balb/c mouse
The Rhipicephalus (Boophilus) microplus is one of the most important arthropods in veterinary medicine due economic losses and health problems caused in cattle production, mainly in Central and South America, as well as in Australia. As an alternative to chemical control, immunization of bovines with Bm86 antigen to induce a protective immune response. A synthetic peptide, SBm 7462, derived from Bm86, has been shown great results in control of ticks. The construction and synthesis of one nucleotide sequence based on this peptide might be useful for design a DNA vaccine that has many advances than peptide vaccine. A gene, called seq1, was constructed with a three repetition of nucleotide sequence of SBm 7462. It was cloned into a pCIneo vector expression in mammals, transfected in VERO cells and injected in BALB/c mouse. Two methods were used to analysis of peptide expression in vitro: DOT?ELISA and PAP. In both, the results showed that the nucleotide sequence (seq1) had not been express in VERO cells. In vivo, when mice were inoculated with the expression cassette they did not response in ELISA. They elevated antibody titles only when vaccinated with the syntetic peptide SBm 7462. And, the best titles of immunoglobulins were seen when the SBm 7462 was administered subcutaneously. After that, we insert a mutation at the begging of seq1, but, the sequenciament demonstrated that any initiation codon (ATG) had been inserted.
Advisor:Sérgio Oliveira de Paula; Márcia Rogéria de Almeida Lamêgo; Joaquín Hernán Patarroyo Salcedo; Marlene Isabel Vargas Viloria; Juliana Lopes Rangel Fietto
School:Universidade Federal de Viçosa
Source Type:Master's Thesis
Date of Publication:05/16/2008