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Figure 1 | Veterinary Research

Figure 1

From: Disruption of the M949_RS01915 gene changed the bacterial lipopolysaccharide pattern, pathogenicity and gene expression of Riemerella anatipestifer

Figure 1

Identification of the mutant strain RA1067. A PCR amplification. M: Takara DL2000 marker; lanes 1–2: R. anatipestifer 16S rRNA was amplified from the WT strain CH3 (lane 1), the mutant strain RA1067 (lane 2), showing a 744-bp fragment of 16S rRNA; lanes 4–5: 644 bp fragment of the erm gene was amplified from the mutant strain RA1067 (lane 5), but not from the WT strain CH3 (lane 4); lanes 7–8: 714-bp fragment of M949_RS01915 gene was amplified from the WT strain CH3 (lane 7), but not from the mutant strain RA1067 (lane 8); lanes 3, 6 and 9 distilled water, as negative controls. B Southern blot analysis of the transposon Tn4351 insertion. Lane M Takara DL15000 marker; Lane 1, 10 μg of pEP4351 digested with XbaI (positive control). Lane 2, 10 μg of chromosomal DNA from mutant strain RA1067 digested with XbaI; Lane 3, 10 μg of chromosomal DNA from the WT strain CH3 digested with XbaI (negative control); each digested sample was resolved on a 0.7% agarose gel, and Southern blot was performed using a TnDIG labeled probe. C Schematic chart of Tn4351 insertion in the RA1067 chromosome. D Real time qPCR analysis. The changes of mRNA were expressed as fold expression and calculated using the comparative CT (2−∆∆CT) method. Data were normalized to the housekeeping gene ldh and expressed as fold changes. Error bars represent standard deviations from three replicates (***, p < 0.001).

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