Difference between revisions of "Part:BBa K3507004"
 
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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.
 
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.
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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).
  
 
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[[File:T--Groningen--Ppta.png|500px|thumb|centre|Figure1. The structure of the pta promoter (Presecan-Siedel, 1999)]]
 
[[File:T--Groningen--Ppta.png|500px|thumb|centre|Figure1. The structure of the pta promoter (Presecan-Siedel, 1999)]]
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<h5>Bibliography</h5>
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<p>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.</p>
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<p>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.</p>

Latest revision as of 14:30, 27 October 2020

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.