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Applications of BBa_K4632008

nirB +dapA - Nutritionally deficient Biosafety device based on diaminopimelic acid (DAP)

E. coli wm3064 is a dapA mutant strain with diaminopimelic acid (DAP) nutritional defect. The dapA gene were ligated to the downstream of pnirB and cloned to pUC18T-mini-Tn7T plasmid, then transformed to wm3064. The growth of ΔdapA/dapA- pUC18T-mini-Tn7T and ΔdapA strains were tested under aerobic or anaerobic conditions. (Figure. 4)The results showed that ΔdapA could only grow in the presence of DAP no matter in aerobic or anaerobic conditions, while ΔdapA/dapA- pUC18T-mini-Tn7T could grow under anaerobic conditions without the supplementation for DAP.

These results(Figure. 1) indicate that this strain can only grow under anaerobic conditions and possesses biological safety. During production, growth can be ensured by adding DAP.

Figure. 1 Results of culture experiment of dapA gene defective strain<p> <p>This work qualitatively confirms that the pnirB element can indeed exhibit specific expression under anaerobic conditions. Furthermore, by utilizing pnirB to drive the essential diaminopimelic acid gene dapA, it restricts bacterial growth to anaerobic environments, such as the gut of the S. invicta. These findings provide a theoretical foundation for the next steps in the development of the biological safety device, specifically the recombination of pnirB into the E. coli chromosome. See more detail in BBa_K4632012[1]

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