Contribution

We registered 5 promoters, 5 secretion signals, and 4 cellulose degrading enzymes as new basic parts. Our newly registered P2069M (BBa_K4317001) and PtrnQ (BBa_K4317004) constitutive promoters can be used in both Bacillus and E. coli. In particular, in Bacillus, these promoters showed higher transcriptional activity than the previously known P43 promoter.

[Figure 1. Confirmation of transcriptional activity of various promoters in E. coli and Bacillus subtilis]

And Pmglv (BBa_K4317002) has been reported as a maltose inducible promoter. We compared this promoter with IPTG inducible. In Bacillus, Pgrac100 (BBa_K4317003) inhibited by LacI was strongly induced by IPTG, but Pmglv was slightly induced by maltose. However, when incubated for a longer period of time, the expression level of mScarlet was similarly observed in the tube without maltose (data not shown). The Pgrac100 promoter works like a constitutive promoter in the absence of Pbla::lacI even if there is lacO downstream. On the other hand, if Pbla::lacI (BBa_K4317016) is present in the plasmid DNA, the activity of Pgrac100 in Bacillus is repressed. P2069M and PtrnQ, which we registered and identified, are constitutive promoters usable in both B. subtilis and E. coli.