Abstract
We present results on the initial definition of proteins synthesized by two isolates of Anaplasma marginale. Bovine erythrocytes infected with A. marginale were radioactively labeled with [35S]methionine or a 3H-amino acid mixture during short-term in vitro culture. The labeled proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This technique revealed protein bands of various apparent molecular weights from less than 14,000 to greater than 200,000. The bands observed represented A. marginale proteins because (i) uninfected erythrocytes from the same animal did not incorporate radioisotope during identical culture conditions, and (ii) the incorporation of radioisotope into proteins during culture of infected erythrocytes was inhibited by tetracycline but not by cycloheximide. The radioactive protein profiles of two different isolates of A. marginale, from Washington and Florida, were compared by two-dimensional gel electrophoresis. About 200 proteins were resolved in each case. Several proteins differed in position when the two-dimensional gel maps were compared, indicating variations in protein structure between the two A. marginale isolates.