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Characterization of recombinant MSP5 Anaplasma marginale Havana isolate
B Corona1, H Machado, M Rodríguez
1National Centre for Animal and Plant Health. Apartado 10, postal address 32700, San José de las Lajas , La Habana , Cuba.
Abstract:
Anaplasma marginale is the causative agent of bovine anaplasmosis, a disease of worldwide economic importance. Major surface proteins (MSPs) are involved in host-pathogen and tick-pathogen interactions and they have been used as markers for the genetic characterization of A. marginale strains and phylogenetic studies. The major surface protein 5 (MSP5) is highly conserved in the genus Anaplasma and in all isolates of A. marginale. The aim of the present work was to carry out the cloning, sequencing and characterization of the recombinant MSP5 Anaplasma marginale Havana isolate. The sequence of the msp5 gene of Anaplasma marginale Havana isolate with a size of 633 pb was determined (Acc. No. AY527217). This gene was cloned into pRSETB vector and expressed in Escherichia coli. The MSP5 protein was recognized by the monoclonal antibody ANAF16C1 and it showed a high similitude percent with the gene sequence described for other Anaplasma marginale isolates. These data are very important for the development of a diagnostic test for A. marginale using the MSP5 recombinant protein.
Insights
Researchers cloned and sequenced the major surface protein 5 (MSP5) from Anaplasma marginale. This highly conserved protein is crucial for developing a diagnostic test for bovine anaplasmosis.
Area of Science:
- Veterinary Microbiology
- Molecular Biology
- Parasitology
Background:
- Anaplasma marginale causes bovine anaplasmosis, a significant economic disease.
- Major surface proteins (MSPs) mediate host-pathogen interactions and are used for strain characterization.
- MSP5 is a highly conserved protein across Anaplasma species and isolates.
Purpose of the Study:
- To clone, sequence, and characterize the recombinant MSP5 from the Anaplasma marginale Havana isolate.
- To assess the potential of MSP5 as a target for diagnostic test development.
Main Methods:
- Sequencing of the msp5 gene from Anaplasma marginale Havana isolate (633 bp).
- Cloning of the msp5 gene into the pRSETB vector.
- Expression of the recombinant MSP5 protein in Escherichia coli.
- Characterization using monoclonal antibody ANAF16C1.
Main Results:
- The msp5 gene sequence (Acc. No. AY527217) was determined.
- Recombinant MSP5 protein was successfully expressed in E. coli.
- The expressed MSP5 protein was recognized by the ANAF16C1 monoclonal antibody.
- High sequence similarity was observed between the Havana isolate MSP5 and other Anaplasma marginale isolates.
Conclusions:
- The MSP5 gene sequence from the Anaplasma marginale Havana isolate was successfully determined and characterized.
- The recombinant MSP5 protein shows promise for developing a diagnostic test for bovine anaplasmosis.
- The conserved nature of MSP5 supports its utility in diagnostics and phylogenetic studies.
