Mutant strains of mycoplasma hyopneumoniae
A technology for mycoplasma hyopneumoniae and mutant strains, applied in the direction of bacterial antigen components, using vectors to introduce foreign genetic material, microorganisms, etc., can solve the problems that have not yet been developed for mycoplasma hyopneumoniae, and the method of transforming mycoplasma hyopneumoniae has not yet been described
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Embodiment 1
[0329] Embodiment 1.-select Mycoplasma hyopneumoniae bacterial strain
[0330] The wild-type strain of M. hyopneumoniae used was strain 6314 corresponding to the virulent M. hyopneumoniae isolate from Amer-Girona-Spain and from Iowa State University (Iowa State University). Strain 232 from the University) (Ames-Iowa-USA).
Embodiment 2
[0331] Example 2.-Construction of replicative plasmids
Embodiment 21
[0332] Example 2.1.-Construction of replicative plasmid pOG
[0333] To obtain a replicating plasmid in M. hyopneumoniae, the oriC region of M. hyopneumoniae amplified by PCR was introduced into plasmid pBluescript II SK. Oligonucleotide 5OriCNotI(ATG CGC GGC CGC TTA TTT ATC AGA AAC AGT TAG) (SEQ ID NO: 9) and 3OriCXbaI (AGT C GG GCC C AG CTT GCG CAT CAT TGG ATG ATG GAT TC) (SEQ ID NO: 10) was used for this amplification and genomic DNA of M. hyopneumoniae strain J was used as template. The resulting 1.96 kb fragment was then digested with restriction enzymes NotI and XbaI. On the other hand, by PCR using the oligonucleotide 5GmORF (ACT G GG ATC C AT GAA TAT AGT TGA AA ATG) (SEQ ID NO: 11) and 3GmApa (GGA T GG GCC C AG CTT GCG CAT CAT TGG) (SEQ ID NO: 12) and use plasmid pIV-T as a template to amplify the gentamicin resistance gene (aac(6')-aph(2")). Then use the corresponding restriction The 1.8 kb PCR product was digested enzymatically. By PCR using ...
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