Difference between revisions of "Part:BBa S04059"
| (6 intermediate revisions by the same user not shown) | |||
| Line 7: | Line 7: | ||
==== Characterization by team Mingdao 2020 ==== | ==== Characterization by team Mingdao 2020 ==== | ||
| − | + | :;ldhp-AQP-RBS-SpxB-Tr-tpxp-KatG-Tr/pSB1C3 ([https://parts.igem.org/Part:BBa_K3376012 BBa_K3376012]) | |
| − | :Catalase converts H2O2 to O2 and H2O preventing cells from oxidative stress. When E. coli was expressing aquaporin (AQP) and pyruvate oxidase (SpxB) to produce H2O2, the cell growth was inhibited remarkably compared to a vector-only control. By adding of catalase (KatG), the growth rate became normal and have similar response to glucose concentrations with the control. | + | :Catalase converts H2O2 to O2 and H2O preventing cells from oxidative stress. When E. coli Nissle strain (EcN) was expressing aquaporin (AQP) and pyruvate oxidase (SpxB) to produce H2O2, the cell growth was inhibited remarkably compared to a vector-only control. By adding of catalase (KatG), the growth rate became normal and have similar response to glucose concentrations with the control. |
[[File:T--Mingdao--ww5.png|600px|center]] | [[File:T--Mingdao--ww5.png|600px|center]] | ||
| + | |||
| + | |||
| + | :The lysates of the transformed E. coli Nissle carrying the indicated gene were subjected to SDS-PAGE and Coomassie blue staining. When overexpression under the promoter of ldhp, KatG (80kDa) and AQP (17kDa) have sharp band on the PAGE. However, the predicted 65-kDa of SpxB was not detected, possibly because of the oxidative stress induced by the production of H2O2. In the presence of KatG, SpxB was shown up along with AQP, though expressed at a week level. | ||
| + | |||
| + | [[File:T--Mingdao--ww10.png|600px|center]] | ||
| + | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
| Line 17: | Line 23: | ||
<!-- --> | <!-- --> | ||
| + | |||
=== <span class='h3bb'>Sequence and Features</span> === | === <span class='h3bb'>Sequence and Features</span> === | ||
<partinfo>BBa_S04059 SequenceAndFeatures</partinfo> | <partinfo>BBa_S04059 SequenceAndFeatures</partinfo> | ||
Latest revision as of 02:10, 15 October 2020
katG (LAA).B0015
Construction intermediate
Characterization by team Mingdao 2020
- ldhp-AQP-RBS-SpxB-Tr-tpxp-KatG-Tr/pSB1C3 (BBa_K3376012)
- Catalase converts H2O2 to O2 and H2O preventing cells from oxidative stress. When E. coli Nissle strain (EcN) was expressing aquaporin (AQP) and pyruvate oxidase (SpxB) to produce H2O2, the cell growth was inhibited remarkably compared to a vector-only control. By adding of catalase (KatG), the growth rate became normal and have similar response to glucose concentrations with the control.
- The lysates of the transformed E. coli Nissle carrying the indicated gene were subjected to SDS-PAGE and Coomassie blue staining. When overexpression under the promoter of ldhp, KatG (80kDa) and AQP (17kDa) have sharp band on the PAGE. However, the predicted 65-kDa of SpxB was not detected, possibly because of the oxidative stress induced by the production of H2O2. In the presence of KatG, SpxB was shown up along with AQP, though expressed at a week level.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 228
Illegal BglII site found at 1163
Illegal BglII site found at 1290
Illegal BamHI site found at 1080 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]