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To test whether gntR (BMEI0106) has any influence on ROCK inhibitors for glaucoma B. melitensis virulence, a gntR (BMEI0106) mutant was constructed and its virulence was evaluated. BALB/c mice were inoculated intraperitoneally with B. melitensis and spleen colonization was assessed at different time point postinfection. Compared to the wild-type strain 16M and the complementary strain 16M��gntR-C, splenic CFU in 16M��gntR infected mice were significantly reduced (Figure ?(Figure4A).4A). The results demonstrated that gntR was important for the intracellular survival of B. melitensis. Additionally, RT-PCR analysis showed gntR was highly activated during mouse infection (Figure ?(Figure4B),4B), again confirming its role in the intracellular survival of B. melitensis. Moreover, the expression profile of gntR in vivo was contrast to BSR0602 (Figures ?(Figures2C2C and ?and4B),4B), confirming BSR0602 negatively regulates BMEI0106. Hence, BSR0602 may modulate B. melitensis intracellular survival through repressing gntR. Figure 4 BSR0602 regulation Brucella virulence by targeting gntR. (A) Contribution of GntR (BMEI0106) to the virulence of B. melitensis. Groups of five BALB/c mice were infected intraperitoneally with 2?��?106?CFU of 16M, 16M��gntR, ... Our results showed that gntR was overproduced in BSR0602 mutant than that in the wild type strain (Figure ?(Figure3A),3A), so we supposed that BSR0602 mutant perhaps be more virulent than the wild type. To determine whether this is the case, the deletion mutant 16M��BSR0602 and the overexpression Non-specific serine/threonine protein kinase Crizotinib strain 16M-BSR0602 were characterized for changes in virulence related phenotypes. First, 16M, 16M��BSR0602, and 16M-BSR0602 were tested for their survival capability under various environmental stress conditions that simulate the intracellular environments encountered by Brucellae during their dissemination. Compared to the wild strain 16M, the survival of BSR0602 mutant 16M��BSR0602 under oxidative stress, high-temperature, and osmotic stress did not change significantly (P?>?0.05) (Figure ?(Figure5A).5A). The survival percentage of 16M��BSR0602 only decreased slightly (12%) upon exposure to acidic pH. However, the overexpression strain 16M-BSR0602 was significantly reduced in its ability to survive under all stress conditions (P?