Gaps Repairing And Analysis Of Mycoplasma Bovis NM 2012 Complete Genome Sequence

Mycoplasma bovis is a pathogen which can cause varieties of diseases, such as peneumonia, arthritis, abortion, mastitis, and so on. Mycoplasma bovis has caused great economic losses in cattle raising industry, and seriously affected the healthy development of cattle raising industry.The complete genome sequence of M. bovis NM 2012 was finished by Wang Yanjie, However, there were sequence gaps and redundant repeat sequences. The objective of this study is to obtain the complete genome sequence of M. bovis NM 2012. M. bovis HB0801 was selected as a reference gene sequences in this test, after compared with M.bovis HB0801, there were 34 unfinished sequence. In order to obtain M.bovis NM 2012 complete genome sequences, we designed primers, performed PCR amplification, cloned and sequenced, then used bioinformatics software to assemble genome sequence and predicted its ORFs、lipoprotein、insertion sequence (IS) elements, and so on. The alignment result showed that there were 21 gaps used N instead of unknown gene sequences in 34 unfinished sequences, and N ranging from 10bp to 2,359bp, a total number of N was 13,131bp.34 unfinished sequences included 23 gaps and 11 redundant repeat sequences, there were 24,763bp in 23 gaps, and 14,765bp in 11 redundant repeat sequences. Cloning sequence results showed that a total of missing sequence were 20,222bp in 23 gaps.11 redundant repeat sequences length of 14,765bp, actually 10,226bp were invalid repeat sequences and 4,539bp were its own sequences. M.bovis NM2012 was sequenced by Wang Yanjie, and the complete genome size of this strain was 993,483bp. In this test, we removed 13,131bp of N and 10,226bp of invalid repeat sequences, then filled 20,222bp of missing sequences, we finally obtained complete genome size of the M.bovis NM 2012 strain was 990,348bp.The complete genome size of the M.bovis NM 2012 strain was 990,348bp, the content of GC% was 29.29%; After alignmented with M.bovis HB0801 and M.bovis NM 2012, we predicted coding region contained 839 ORFs、34 tRNA、6 rRNA、103 lipoproteins and 2 Vsp. Based on the 5 strains of Mycoplasma bovis compared, we can concluded that M.bovis NM 2012 and M.bovis PG45 has smallest similarity, M.bovis NM 2012 and M.bovis HB080I has highest similarity.