[Construction of eag deletion mutant of Bacillus anthracis vaccine strain A16R].

OBJECTIVE

Construction of eag deletion mutant of Bacillus anthracis vaccine strain A16R.

METHODS

To study the function of the gene eag of Bacillus anthracis vaccine strain A16R, according to the sequence of Bacillus anthracis Ames strain, we designed primers and constructed a recombinant plasmid by the spectinomycin resistance cassette, upstream homologous fragment and downstream homologous fragment of eag cloned in tandem in pKSV7. We introduced the recombinant into A16R by electroporation and screened the mutant using the principle of homologous recombination. We checked the mutant using the PCR and proteomics.

RESULTS

We constructed the recombinant plasmid successfully and got the eag deletion mutant. PCR results showed the gene eag was deleted; SDS PAGE showed evident differences between prime strain and mutant strain. Two-dimensional gel electrophoresis results displayed three EA1 protein points of prime strain were absent in the mutant strain.

CONCLUSION

We constructed eag deletion mutant of Bacillus anthracis vaccine strain A16R. This research will be helpful to study the functions of eag gene and the other important genes of Bacillus anthracis.