Abstract:

To express P97-R1 gene on Z strains of Mycoplasma hyopneumoniae for the exploration of possibility of use as a diagnostic antigen, a pair of primers were designed according to the online database of P97 genes of Mhp232 on GenBank, a strain of Mycoplasma hyopneumoniae, and subsequently contributed to the successful amplification of interest segments using the genomic DNA of Z strains of Mhp as templates by PCR. P97-R1 gene was ligated with T-easy, sequenced and was used to transform competent E. coli Rosetta .The results was compared with 232 referential strains. It was revealed that there were many sites of mutations in the P97-R1. There were 11 5-aa repeats in that of the Z strain ,15 5-aa repeats in the P97-R1 region of the 232 strains as analyzed by DNAstar software. Speculated that differences in repeat sequence R1 may be the reasons of the strain was strong or weak. Synthesized products were analyzed using SDS-PAGE and 30 KD proteins of the P97-R1 was detected. The reactionogenicity was then verified by Western-blotting. This work laid a foundation for the diagnosis, prevention Mhp and further research the Mycoplasma hyopneumoniae.