Isolation And Identification Of Mycoplasma Bovis And Analysis Of Its Complete Genome Sequence

This test isolated a suspected mycoplasma strains from pneumonia and arthritis cases of calves, initially identified as Mycoplasma bovis by pathogen isolation, culture and biochemical identification method. Identified as Mycoplasma bovis by PCR amplified specific oppD/F fragment of Mycoplasma bovis and sequenced PCR products and blasted.This strains was named as Mycoplasma bovis NM2012. To understand the whole genome information of NM2012, the complete genome sequence of this strains were sequencing and analysis, lay the data foundation for further study of the strains.The results were as follows:The strains can form smooth, white small colony on mycoplasma solid medium and "fied eggs" colony morphology under the microscope, it has center umbilical.The resuLts of biochemical identification shows that the strains cannot ferment glucose, hydrolyze arginine, did not decompose urea and lique faction of gelatin was negative.The strains specific PCR fragments consistent with the purpose fragment and99%homology with Mycoplasma bovis after blast.The complete genome size of this strains was993,483bp, the content of GC%was29.26%.The strains contained885ORFs after predicted,the total length was834,042bp,representing83.95%of the full genome,GC%content was29.68%in ORFs region.The total length of intergenic were159,441bp,accounting for16.05%of the genome,GC%content was27.06%in intergenic region.The number of tandem repeats were79,the total length were9,203bp, accounting for0.92%of the total length of the genome, the number of satellites and microsatellite were53and5.The number of tRNA and rRNA were34and4.Analysis of this strains found that10genes were associated with variable surface lipoprotein, this strains had gene for encoding enolase and chaperone protein DnaK, DnaJ and GrpE.34tRNA encoded21kinds of amino acids.4rRNA was identificad in this strains including5S rRNA,5S rRNA,16S rRNA and23S rRNA.