H-NS Represses Biofilm Formation and c-di-GMP Synthesis in Vibrio parahaemolyticus.

Objective: This study aimed to investigate the regulation of histone-like nucleoid structuring protein (H-NS) on biofilm formation and cyclic diguanylate (c-di-GMP) synthesis in Vibrio parahaemolyticus RIMD2210633. Methods: Regulatory mechanisms were analyzed by the combined utilization of crystal violet staining, quantification of c-di-GMP, quantitative real-time polymerase chain reaction, LacZ fusion, and electrophoretic-mobility shift assay. Results: The deletion of hns enhanced the biofilm formation and intracellular c-di-GMP levels in V. parahaemolyticus RIMD2210633. H-NS can bind the upstream promoter-proximal DNA regions of scrA, scrG, VP0117, VPA0198, VPA1176, VP0699, and VP2979 to repress their transcription. These genes encode a group of proteins with GGDEF and/or EAL domains associated with c-di-GMP metabolism. Conclusion: One of the mechanisms by which H-NS represses the biofilm formation by V. parahaemolyticus RIMD2210633 may be via repression of the production of intracellular c-di-GMP.

[The preparation of rabbit antiserum against gekko japonicus Hoxc10 and it's property identification].

AIM: Prepare the rabbit antiserum against gecko japonicus Hoxc10 and to identify its properties. METHODS: Prokaryotic expression vector of g-Hoxc10 were constructed and then transform into E.coli (BL21). To make GST-g-Hoxc10 fusion protein in E.coli (BL21) under the optimized induction of Isopropyl ß-D-1-thiogalactopyranoside(IPTG). The recombination proteins were purified using affinity chromatography. The purified fusion protein was inoculated into adult rabbits to develop antiserum. Western blot and immunohistochemistry staining were then performed to evaluate the feature of the prepared antiserum. RESULTS: Prokaryotic expression vectors of g-Hoxc10 were successfully constructed. The soluble recombinant protein was highly expressed in E.coli BL21 and inoculated into adult rabbits to obtain high titer antiserum. Western blot and immunohistochemistry staining were then performed to evaluate the specificity of the prepared antiserum. CONCLUSION: We successfully amplified and expressed the g-Hoxc10 in E.coli BL21. The purified fusion protein was inoculated into adult rabbits to develop antiserum. The obtained antiserum of g-Hoxc10 showed a high titer against Hoxc10 proteins. The protein and antiserum prepared in this study can be used for further research of the function investigation of Hoxc10.