Difference between revisions of "Part:BBa K3972004"
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[[File: T--TU-Eindhoven--combiB ARG.jpeg|200px|]] | [[File: T--TU-Eindhoven--combiB ARG.jpeg|200px|]] | ||
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''Figure 2. Agar plate with transfected ARG1 with the insert in XL10-Gold cells.'' | ''Figure 2. Agar plate with transfected ARG1 with the insert in XL10-Gold cells.'' | ||
[[File:T—TU_Eindhoven--combiB.png|200px|]] | [[File:T—TU_Eindhoven--combiB.png|200px|]] | ||
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''Figure 3. Agar plate with no cotransfected ARG1, TtrR and TtrS in BL21 (DE3).'' | ''Figure 3. Agar plate with no cotransfected ARG1, TtrR and TtrS in BL21 (DE3).'' | ||
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==Sequence and Features== | ==Sequence and Features== |
Revision as of 11:58, 15 October 2021
Tetrathionate sensor with transferred TtrR and pTtrB185-269 activated ARG1
This composite part is a combination of the parts BBa_K3972001 and BBa_K3972002, with additional functional parts BBa_R0040, BBa_K3332034 and BBa_K2507019. The ARG1 proteins are controlled by the pTtrB185-269 promoter. This promoter is activated by TtrR in the presence of TtrS and tetrathionate. In the presence of tetrathionate, TtrS is activated and in its turn TtrS activates TtrR. TtrR binds to the pTtrB185-269 promoter and induces expression of the ARG1 gas vesicle proteins
Figure 1. The mechanism of the composite part tetrathionate sensor with transferred TtrR and pTtrB activated ARG1.
Usage and Biology
This part is optimized for expression of ARG1 in E. coli cells. This part was successfully transformed after multiple attempts in XL10-Gold and Top10 cells. The cotransformation together with TtrS did also result in colonies on plates with BL21 (DE3), Top10 and XL10-Gold cells. Unfortunately, there was no time left for testing this part with ultrasound equipment.
Characterization
Expression
This part is optimized for expression of ARG1 in E. coli cells. This part was successfully transformed after multiple attempts in XL10-Gold and Top10 cells. The cotransformation together with TtrS did also result in colonies on plates with BL21 (DE3), Top10 and XL10-Gold cells. Unfortunately, there was no time left for testing this part with ultrasound equipment.
Figure 2. Agar plate with transfected ARG1 with the insert in XL10-Gold cells.
Figure 3. Agar plate with no cotransfected ARG1, TtrR and TtrS in BL21 (DE3).
Sequence and Features
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1658
Illegal EcoRI site found at 4811
Illegal XbaI site found at 1409
Illegal PstI site found at 404
Illegal PstI site found at 4668
Illegal PstI site found at 4773
Illegal PstI site found at 5151 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1658
Illegal EcoRI site found at 4811
Illegal NheI site found at 1543
Illegal PstI site found at 404
Illegal PstI site found at 4668
Illegal PstI site found at 4773
Illegal PstI site found at 5151 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1658
Illegal EcoRI site found at 4811
Illegal BglII site found at 2162
Illegal BamHI site found at 5567
Illegal XhoI site found at 5255 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1658
Illegal EcoRI site found at 4811
Illegal XbaI site found at 1409
Illegal PstI site found at 404
Illegal PstI site found at 4668
Illegal PstI site found at 4773
Illegal PstI site found at 5151 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1658
Illegal EcoRI site found at 4811
Illegal XbaI site found at 1409
Illegal PstI site found at 404
Illegal PstI site found at 4668
Illegal PstI site found at 4773
Illegal PstI site found at 5151
Illegal NgoMIV site found at 2613
Illegal NgoMIV site found at 3015
Illegal AgeI site found at 334
Illegal AgeI site found at 5534 - 1000COMPATIBLE WITH RFC[1000]