Cyclic diguanylate (c-di-GMP) regulates the motility of Leptospira interrogans via chemotaxis regulatory proteins CheB/CheR
10.3760/cma.j.issn.0254-5101.2017.07.008
- VernacularTitle:c-di-GMP通过趋化调节蛋白CheB/CheR调控钩端螺旋体运动
- Author:
Liangliang KONG
;
Guohui XIAO
;
Fudi YUAN
;
Jie YAN
;
Xu′ai LIN
;
- Keywords:
Leptospira interrogans;
Chemotaxis;
CheB;
CheR;
c-di-GMP
- From:
Chinese Journal of Microbiology and Immunology
2017;37(7):521-526
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the regulatory effects of cyclic diguanylate (c-di-GMP) signaling on CheB and CheR, which were chemotaxis regulatory proteins relating to the motility of Leptospira interrogans.Methods Real-time PCR was used to determine the expression of cheB1, cheB2, cheB3, cheR1 and cheR2 genes at mRNA level during Leptospira interrogans infection.Fragments of these genes were amplified and cloned into the expression vector pET-28a, respectively, to construct the prokaryotic expression system for them.Colony morphologies of Escherichia coli (E.coli) strains that overexpressed the target genes were observed to determine the regulatory effects of c-di-GMP on CheB and CheR.Results The expression of cheB1 gene at mRNA level increased 60 min after infection and reached the peak at 90 min.Compared with the control group, the expression of cheB3 gene at mRNA level were up-regulated, while no significant difference in the expression of cheB2 and cheR genes was observed 60 min after infection.The prokaryotic expression system for the five genes was successfully constructed and the purified proteins were obtained.CheB1, CheB3 and CheR2 improved the motility of E.coli, but that was inhibited by the inhibitor of diguanylate cyclase (DGC) or phosphodiesterase (PDE).Conclusion CheB and CheR regulate the swarming motility of E.coli and are affected by intracellular c-di-GMP.
