Fig. 4
RhsPi (VP1518) is the immunity protein to RhsP. a RhsPi from V. parahaemolyticus RIMD2210633, not VP1519 or VP1520, neutralized the toxicity of WHH domain in E. coli DH5α. WHH domain was expressed under the control of arabinose and other tested genes were expressed under the control of IPTG in E. coli DH5α. b Killing assays between various strains of V. parahaemolyticus. Prey cells contain a pBBR332 plasmid (kanamycin resistance) for selection purpose. The means of each data set in one representative experiment performed with quadruplicates were shown. Equivalent results were obtained at least three times. Asterisks mark statistical significance of bacterial CFU between sample groups at t = 4 h (P < 0.05) by the two-tailed Student t test. N.S., no significant difference (P > 0.01). c RhsPi binds to RhsPC. Myc epitope-tagged RhsPi (RhsPi-M) was immuno-precipitated by FLAG-tagged RhsP (RhsPC-F). Additional VSV-G tagged PAAR2 (PAAR2-V) broke their interactions. RhsPC-F (pBAD24-rhsPC-F), RhsPi-M (pCX340-rhsPi-M), or PAAR2-V (pBAD24-paar2-V) was expressed in E. coli DH5α. The ratio of PAAR2 to RhsPi (0.5:1, 1:1, and 2:1) shown in the figure represents the original amount of bacteria expressing respective protein used for lysis and the subsequent mixture for Co-IP
