MOLECULAR CHARACTERIZATION OF MEMBRANE PROTEIN GENES OF ANAPLASMA MARGINALE FROM RIO DE JANEIRO STATE, BRAZIL
Autor(es): Pedro Bittencourt , Huarrisson Azevedo Santos, Ananda Müller, Danilo Fonseca Balbi, Carlos Luiz Massard, Nadia Regina Pereira Almosny |
| Anaplasma marginale causes illnesses and economic losses and it’s considered one the most frequent and important organisms infecting cattle in Brazil. Since vaccination is the main strategy for the control of A. marginale, currently the search for new antigens able to increase the level of protection and/or even prevent the carrier state is considered of great importance. The sequencing of the complete genome of A. marginale has made it possible to identify several genes that encoding membrane proteins. However, there is little knowledge regarding the genetic variability of those genes in geographically distinct A. marginale isolates. This study evaluated the degree of conservation between the predictive amino acid sequences of omp1, omp4, omp5, omp7 , SodB, am097, AM254 (ef-tu) and am956 (PEPA) genes in A. marginale samples from five municipalities representing four regions of Rio de Janeiro State. After polimerase chain reaction amplification, the products Sanger method and the predicted amino acid sequence were aligned using the MEGA 4 software, comparing values of the number of amino acid differences per site ("p-distance") between the predicted sequences of the five RJ samples and the Pernambuco-Zona da Mata, Saint Maries, Florida isolates of A. marginale. The analysis of the mean between all predictive amino acid sequences evaluated indicates that there is little variability in this protein. According to these data, EF-Tu, SODb, PEPa, and OMP1 OMP4 proteins are less variable, with values of "p-distance" of 0.01, 0.02, 0.02, 0.02 and 0 02 respectively. Only OMP7 and VirB9-1 showed values higher than 5%, 0.11 and 0.08, respectively. The samples of Rio de Janeiro State showed greater similarity in predictive sequences among themselves than when compared with the other samples from both Brazil and the USA, suggesting that the use of these proteins as potential immunogens may be limited to each specific region. |
