Difference between revisions of "Part:BBa K4385019"
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<partinfo>BBa_K4385019 short</partinfo> | <partinfo>BBa_K4385019 short</partinfo> | ||
− | + | This part is composed of CusSR two-component signal transport system, PcusC and OmpC-CBP. | |
− | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
+ | Promoter PCusC can express downstream genes under the induction of Cu2+. When the zinc ion concentration in the environment exceeds a certain concentration, the PcusC promotes the transcription of downstream OmpC-CBP, and realizes the adsorption of zinc ions in the environment. The 6*His Tag was added to this sequence to purify the recombinant protein. | ||
+ | ===Characterization=== | ||
+ | Therefore, PCusC-OmpC-CBP-ST was constructed so that the OmpC-CBP would be expressed only the presence of copper ions in the environment. | ||
+ | [[Image: Cu_isd.png|center|frame|100px|<b>Figure 1.PCusC-OmpC-CBP-ST.</b>]]<br><br> | ||
+ | |||
+ | To verify the construction of the PCusC-OmpC-CBP-ST, the enzyme digestion and agarose gel electrophoresis were performed (Fig. 2A). | ||
+ | After 4 hrs of induction with 0.5, 1.0, 2.0 mM CuSO4 respectively, the concentration of Cu2+ in the supernatant was measured. The result also shows that our engineered bacteria have better capacity of copper absorption (Fig. 2B) | ||
+ | [[Image: Cu_isd2.png|center|frame|100px|<b>Figure 2. Validation of the absorption of copper ions with PCusC.</b>(A) Digestion and gel electrophoresis of PCusC-OmpC-CBP-ST. (B) The measurement of the concentration of Cu2+ in the supernatant. The Cu2+ was added when the OD600 value of bacteria reached 0.5. The concentration in supernatant was measured after 4 hrs of induction and normalized by the same OD600 value. *,P<0.05 from control group. △,P<0.05 from respective control.]]<br><br> | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> |
Latest revision as of 04:05, 12 October 2022
Copper-inducible surface display
This part is composed of CusSR two-component signal transport system, PcusC and OmpC-CBP.
Usage and Biology
Promoter PCusC can express downstream genes under the induction of Cu2+. When the zinc ion concentration in the environment exceeds a certain concentration, the PcusC promotes the transcription of downstream OmpC-CBP, and realizes the adsorption of zinc ions in the environment. The 6*His Tag was added to this sequence to purify the recombinant protein.
Characterization
Therefore, PCusC-OmpC-CBP-ST was constructed so that the OmpC-CBP would be expressed only the presence of copper ions in the environment.
To verify the construction of the PCusC-OmpC-CBP-ST, the enzyme digestion and agarose gel electrophoresis were performed (Fig. 2A). After 4 hrs of induction with 0.5, 1.0, 2.0 mM CuSO4 respectively, the concentration of Cu2+ in the supernatant was measured. The result also shows that our engineered bacteria have better capacity of copper absorption (Fig. 2B)
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 535
Illegal XbaI site found at 28
Illegal SpeI site found at 694
Illegal PstI site found at 1042 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 535
Illegal NheI site found at 1410
Illegal NheI site found at 1433
Illegal SpeI site found at 694
Illegal PstI site found at 1042 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 535
Illegal BglII site found at 243
Illegal BglII site found at 1545 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 535
Illegal XbaI site found at 28
Illegal SpeI site found at 694
Illegal PstI site found at 1042 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 535
Illegal XbaI site found at 28
Illegal SpeI site found at 694
Illegal PstI site found at 1042
Illegal AgeI site found at 361 - 1000COMPATIBLE WITH RFC[1000]