Clonagem e expressão de epítopos da proteína 1 maior de superfície de Anaplasma marginale em Escherichia coli e Bacillus subtilis

Abstract

Bovine Anaplasmosis is a disease that affects ruminants, especially cattle in Brazil generating economic losses. Among the clinical signs, fever are present in high peaks of parasitemia, as well as anemia, which can lead the animal to death. Ways to combat the disease on the market are not efficient. Animals immunized with surface proteins of Anaplasma marginale (MSPs) showed the humoral immune response, and immunodominant epitopes of MSP1α has being proven an excellent alternative to adaptive immunity. Since immunity to A. marginale is associated with these proteins, which are exposed on the surface rickettsia and are easily accessible by the host immune system, can be neutralized by antibodies against epitopes exposed. In this study, the objective was to express immunogenic epitopes of MSP1α protein in E. coli and Bacillus subtilis. Methods: In this way, two proteins (q1ANAP and q2ANAP) were designed with surface protein epitopes of A. marginale. Results and Conclusion: Purification and protein expression in E. coli with q2ANAP-pRSET vector showed satisfactory results demonstrating that the protein is soluble, whereas in Bacillus subtilis no expression was obtained. The validation of the protein was performed with sera from animals infected with the disease. Thus, we can conclude that q2ANAP has immunogenic epitopes and further studies can be realized aiming to evaluate the protective immune response of infected cattle.