Cloning,expression and immunological characterization of P30 protein of Mycoplasma ovipneumoniae Y98

According to the Mycoplasma hyopneumoniae(Mhp) transmembrane protein P30 gene sequence,a pair of primers was designed,and the P30 gene of Mycoplasma ovipneumonia(MO) membrane protein was obtained by PCR.The result of the homology analysis showed that the MO P30 gene shares 79% identity with the Mhp P30 gene.The results of predictions of the epitopes and the transmembrane of the MO P30 showed that both of transmembrane and epitope were high identity.The fragment contains a TGA which coded Trp rather than the stop codon.The P30 gene was constructed in the prokaryotic expression vector pET-28a and the plasmid was transformed into the strain Rossta to obtain the recombinant strain Rossta(pET-28a-P30).SDS-PAGE and Western blot was performed,the result indicated one kind of 30 000 protein was induced and this protein was mainly in the form of inclusion bodies.Finally,immune experiments showed that prokaryotic expression P30 proteins has a certain protective effect.