[Construction and identification of virB2 deletion mutants of Brucella vaccine strain M5-90].

OBJECTIVE

To construct Brucella vaccine strain M5-90 delta virB2 mutants.

METHODS

Suicide plasmid pGEM-7zf-delta virB2-sacB was constructed by traditional molecular biology technology. Through the method of homologous recombination, we screened Brucella gene deletion mutant by 100 mg/L ampicillin resistance screening and 5% sugar sensitivity screening after electroporation. The M5-90 delta virB2 mutant was identified by PCR. Its stability was detected by continuous bacteria culture.

RESULTS

The virB2 deletion mutant strain was successfully constructed. The reverse mutation did not occur within 10 passages.

CONCLUSION

The results of this study will be based on developing the new attenuated vaccine of Brucella gene deletion mutants.