[Expression of Edwardsiella tarda luxS gene at different growth stage].

ABSTRACT

OBJECTIVE: Purpose of this work was to explore the distribution of LuxS/AI-2 quorum sensing system in Edwardsiella, and analyze expression characteristics and biological function of the key gene luxS accompanying the growth of Edwardsiella. METHODS: The full-length of AI-2/LuxS of Edwardsiella tarda was cloned by PCR based on the sequence on NCBI, then characteristics and conservative structure of this protein-coding gene were analyzed using web database and bioinformatics tools. The anti-rabbits serum was prepared after this protein was purified through prokaryotic expression. The expression level of luxS gene was analyzed during different growth stages using Western blot and further the distribution of luxS gene in Edwardsiella tarda was studied by this technique. To explore whether the specific LuxS is AI-2 dependent we used the method of antibody neutralization to analyze the effect of the anti-rabbits serum on the growth of Edwardsiella tarda. RESULTS] The luxS gene was obtained by PCR, its length was 516 bp, and the sequence was highly conserved in Edwardsiella tarda. Results of Western blot analysis showed that LuxS expression level was the lowest in the lag phase and began increasing when entered index phase. It reached the peak in the late index phase and decreased in decline phase. Moreover, Antibody neutralization results showed that, it can elongate the growth plateau phase, but it has no significant effect on bacterial growth. CONCLUSION: The key gene of luxS was highly conserved, and LuxS/AI-2 was widely distributed among Edwardsiella tarda. The expression level of luxS gene was different during every growth period, expression of LuxS protein reached the highest level in the late index phase.