Construction of cheA insertion mutant of Campylobacter jejuni and the alteration in coloning of mice

Abstract:

Abstract：[Objective] To construct cheA (chemotaxis, che) insertion mutant of Campylobacter jejuni and to observe the role cheA gene plays in coloning mice of Campylobacter jejuni. [Methods] we generated cheA gene insertion mutant of C. jejuni NCTC11168 based on homologous recombination. The cheA mutant was checked by PCR and sequencing. We detected the difference in coloning mice between cheA mutant and wild-type of C. jejuni NCTC11168 by CFU(Colony-Forming Units) counting of C. jejuni in jejunal content. We conformed the role cheA gene plays in coloning mice of Campylobacter jejuni by conplementation analysis. [Results] PCR results reveal that we have successfully construted cheA insertion mutant of C. jejuni NCTC11168. The cheA mutant displayed significantly attenuated colonization on jejunal mucosa of mice compared to wild-type strain (P<0.05). Complementation analysis shows that the complementation of cheA mutant regained its ability in coloning on jejunal mucosa of mice. [Conclusion] The cheA mutant and it’s complementation were successfully construted in our research. The cheA gene may play an important role in coloning of C. jejuni on jejunal mucosa of mice.

Key words:Keywords: Campylobacter jejuni; homologous recombination; chemotaxis