Prokaryotic Expression And Immunological Characteristics Analysis Of The Hemolysin Protein From Mycoplasma Ovipneumoniae

Mycoplasma ovipneumoniae is a highly contagious disease that leads to Mycoplasma pneumonia in sheep and goats spreading all the world. Due to characteristics of mycoplasma, It is not effective by antibiotics therapy. Because it is difficult to culture and has long growth cycle, the traditional vaccine has some problems that hard to overcome. Then we begin to develop new type vaccine with the development of biological chemistry and Biology Information Technology.This study shows it has hemolysis crisis during cultring mycoplasma. The hemolysin protein gene was cloned, the activity of protein was discussed. Fristly, TGA is termination codon in gene cloning, that mutated into TGG by site-specific mutagenesis with consistency in mycoplasma system. We constructed the protein expression systems and mature peptide portion of protein in Escherichia coli. SDS-PAGE showed that the molecular weight was44.5kDa, the fusion protein was immunogenic by Western blot analysis.Mice were immunized four times with purified fusion proteins. The control groups were injected with PBS. The serum antibody titers of vaccinated mice were tested per seven days. The results show the serum antibody titers of immunization group were highly significantly different from that of the control group(P<0.05). After four times immunizations, the results of T lymphocyte Proliferation assay revealed that T lymphocyte Proliferation ability of fusion proteins immunization groups were highly significantly different from that of the control group(P<0.05)...