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. 2017 Mar 3;7:44. doi: 10.1038/s41598-017-00121-y

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© The Author(s) 2017

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Adhesion of M. bovis ^NOX− to EBL cells. (A) The result of sequencing Tn4001. The terminal sequence of Tn4001 was used for the amplification of sequence next to it. Then the sequence was used to map transposon insertion site and confirmed the NOX gene was inserted at 1276 nt (C-terminal of NOX gene). (B) The expression of NOX in M. bovis ^NOX−. The total proteins of M. bovis ^NOX− and M. bovis ^WT were incubated with antiserum against rPGK (1:500) or mAb (1:1000) to rNOX to detect the expression of NOX gene. The cropped blots are displayed here, the full-length blots are presented in supplementary information (Fig. S5). (C) Image J software was used to calculate the NOX/PGK ratio and then the expression of disrupted nox gene in M. bovis ^NOX− was compared with that intact nox in M. bovis ^WT. (D) The adhesion rates of M. bovis ^NOX− at three time points with MOI of 1:1000. ***p < 0.001 represents very significant difference.