Optical mapping of the Mycobacterium avium subspecies paratuberculosis genome.
BACKGROUND
Infection of cattle with Mycobacterium avium subspecies paratuberculosis (M. ap) causes severe economic losses to the dairy industry in the USA and worldwide. In an effort to better examine diversity among M. ap strains, we used optical mapping to profile genomic variations between strains of M. ap K-10 (sequenced strain) and M. ap ATCC 19698 (type strain).


RESULTS
The assembled physical restriction map of M. ap ATCC 19698 showed a genome size of 4,839 kb compared to the sequenced K-10 genome of 4,830 kb. Interestingly, alignment of the optical map of the M. ap ATCC 19698 genome to the complete M. ap K-10 genome sequence revealed a 648-kb inversion around the origin of replication. However, Southern blotting, PCR amplification and sequencing analyses of the inverted region revealed that the genome of M. ap K-10 differs from the published sequence in the region starting from 4,197,080 bp to 11,150 bp, spanning the origin of replication. Additionally, two new copies of the coding sequences > 99.8% were identified, identical to the MAP0849c and MAP0850c genes located immediately downstream of the MAP3758c gene.


CONCLUSIONS
The optical map of M. ap ATCC 19698 clearly indicated the miss-assembly of the sequenced genome of M. ap K-10. Moreover, it identified 2 new genes in M. ap K-10 genome. This analysis strongly advocates for the utility of physical mapping protocols to complement genome sequencing projects.
Wu
Chia-wei
CW
The Laboratory of Bacterial Genomics, Department of Pathobiological Sciences, University of Wisconsin-Madison, WI, USA. chiaweiwu@wisc.edu
Schramm
Timothy M
TM
Zhou
Shiguo
S
Schwartz
David C
DC
Talaat
Adel M
AM
D002874:Chromosome Mapping / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D014644:Genetic Variation; D016680:Genome, Bacterial; D016927:Mycobacterium avium subsp. paratuberculosis / Q000145:classification / Q000235:genetics*; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction
D004269:DNA, Bacterial