Mycoplasma gallisepticum has three virulent genes (mgc1, mgc2 and mgc3), also Mycoplasma synoviae has adhesion gene (vlhA). These genes are responsible for adhesion and colonization of the respiratory tract of the host. Therefore, this study was concerned with preparation of recombinant vaccine from field isolates and compared it with membrane and whole cells vaccines. The virulence genes were cloned into plasmid vectors followed by transformation into E. coli BL-21competent cells for the expression of adhesion proteins. A laboratory experiment was done to evaluate the prepared vaccines. For Mycoplasma gallisepticum recombinant vaccinated vaccinated groups. ELISA results for Mycoplasma recombinant vaccines revealed that there was significant difference between the vaccinated and non-vaccinated vaccinated groups at p < 0.05 for serum and Egg yolk. In the other hand results for ELISA of Mycoplasma synoviae had no significant difference between the means of GMTs of the vaccinated vaccinated groups but overall vaccinated vaccinated groups had a significant high antibody titers than the un-vaccinated vaccinated group at p < 0.05 in both serum and Egg yolk. Concerning polymerase chain reaction and culture, recombinant vaccinated vaccinated group were control from first week post-challenge and continued control till the end of experiment. While membrane and whole cell vaccinated vaccinated groups gave weak positive results with M. gallisepticum polymerase chain reaction and positive for culture (20- 60%).On the other hand, M. synoviae gave positive results (20%) by culture at one and three weeks post challenge and gave weak positive results polymerase chain reaction till the end of experiment. The results showed that the recombinant vaccine gave satisfactory results, than membrane and whole cell bacterins.