Difference between revisions of "Part:BBa R0085"

Revision as of 11:30, 10 October 2022

T7 Consensus Promoter Sequence

The T7 promoter should only produce PoPS when the T7 polymerase is also being expressed.

Literature Characterization by AFCM-Egypt 2022 team

In this study, in comparison to the cytoplasmically-expressing strain, the periplasmic-secreting strain showed faster organophosphorus OPH biocatalytic reaction rates. Additionally, the bacteria under T7 promoter regulation showed faster biocatalytic reaction rates than the strains under Trc promoter regulation, irrespective of the location of OPH inside the cell. The periplasmic-secreting strain's biocatalytic rate was substantially higher with substrate concentrations than the cytoplasmic-expressing strain and showed a 2-fold better conversion rate with 1 mM Paraoxon as shown in figure 2.

Figure 2. bio catalytic activity of E.coli whole cell biosensor under T7 and Trc promoters.

Experimental Characterization by AFCM-Egypt 2022 team

This figure shows an experimental characterization of this part as it's validated through gel electrophoresis as it is in lane 6 (the last one). The run part (ordered from IDT) included T7P - TyrR RBS - TyrR - TyrPromoter.

Usage and Biology

This promoter is the consensus T7 promoter running from -17 to +6.

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]

References

1. Kang, D. G., Seo, S. H., Choi, S. S., & Cha, H. J. (2006). Comparison of whole cell biocatalytic reaction kinetics for recombinant Escherichia coli with periplasmic-secreting or cytoplasmic-expressing organophosphorus hydrolase. Studies in surface science and catalysis, 173-176.

Improvement of R0085 by ZJU-CHINA 2018 Teams

The ZJU-CHINA 2018 Teams fulfilled the improvement of R0085 by inducing some mutation in the sequence. A series of promoters with increased expression strength are constructed. The result can be seen by clicking the link below.

@@ Line 5: / Line 5: @@
 The T7 promoter should only produce PoPS when the T7 polymerase is also being expressed.
-===Usage and Biology===
-This promoter is the consensus T7 promoter running from -17 to +6.
-<span class='h3bb'>Sequence and Features</span>
-<partinfo>BBa_R0085 SequenceAndFeatures</partinfo>
 ==Literature Characterization by AFCM-Egypt 2022 team==
 In this study, in comparison to the cytoplasmically-expressing strain, the periplasmic-secreting strain showed faster organophosphorus OPH biocatalytic reaction rates. Additionally, the bacteria under T7 promoter regulation showed faster biocatalytic reaction rates than the strains under Trc promoter regulation, irrespective of the location of OPH inside the cell. The periplasmic-secreting strain's biocatalytic rate was substantially higher with substrate concentrations than the cytoplasmic-expressing strain and showed a 2-fold better conversion rate with 1 mM Paraoxon as shown in figure 2.
@@ Line 21: / Line 14: @@
 This figure shows an experimental characterization of this part as it's validated through gel electrophoresis as it is in lane 6 (the last one). The run part (ordered from IDT) included T7P - TyrR RBS - TyrR - TyrPromoter.
 <br><br><br><br><br><br><br><br>
+===Usage and Biology===
+This promoter is the consensus T7 promoter running from -17 to +6.
+<span class='h3bb'>Sequence and Features</span>
+<partinfo>BBa_R0085 SequenceAndFeatures</partinfo>