Generator

Part:BBa_K3507004

Designed by: Andreea Stan   Group: iGEM20_Groningen   (2020-10-26)

Constitutive promoter of B. subtilis

The pta promoter of Bacillus subtilis is the region that initiates the transcription of the phosphotransacetylase coding gene. A study by Presecan-Siedel and colleagues from 1999 characterised in detail the pta gene and its promoter from the wild type strain of B. subtilis 168. Fig. 1 taken from this study, illustrates the topology of this promoter as well as the beginning region of the pta gene. Here, they highlighted the putative -10 and -35 regions important for RNA-polymerase binding. For testing the effect of these regions, the up mentioned group deleted the -10 region of the promoter and fused the remaining part to a reporter gene (lacZ). Afterwards, they integrated this fusion at the amyE gene locus of B. subtilis and observed a lower expression level of this genes compared to the expression level when both regions are present. Thus, the promoter is described as a σA –dependent promoter promoter. Another important characteristic for this promoter is the presence of a CRE sequence located between positions -62 and -49 that is responsible for catabolite-activation of this gene. This promoter has recently been used to express GFP in Bacillus mycoides and it showed a to behave as a strong promoter also for this species (Yi et al., 2018).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Figure1. The structure of the pta promoter (Presecan-Siedel, 1999)
Bibliography

1. Presecan-Siedel E, Galinier A, Longin R, Deutscher J, Danchin A, Glaser P, et al. Catabolite regulation of the pta gene as part of carbon flow pathways in Bacillus subtilis. J Bacteriol. 1999;181(22):6889–97.

2. Yi Y, Li Z, Song C, Kuipers OP. Exploring plant-microbe interactions of the rhizobacteria Bacillus subtilis and Bacillus mycoides by use of the CRISPR-Cas9 system. Environ Microbiol. 2018;20(12):4245–60.

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