Difference between revisions of "Part:BBa K4385025"
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<partinfo>BBa_K4385025 short</partinfo> | <partinfo>BBa_K4385025 short</partinfo> | ||
− | This component is made up of TurboID and PbrR. TurboID, a pair of biotin ligases, was designed to overcome the problem of slow existing biotin ligases.PbrR is a lead ion adsorption | + | This component is made up of TurboID and PbrR. TurboID, a pair of biotin ligases, was designed to overcome the problem of slow existing biotin ligases.PbrR is a lead ion adsorption protein. |
− | |||
===Usage and Biology=== | ===Usage and Biology=== | ||
+ | The PbrR without absorbing lead ions was connected and formed a complex under TurbolD's traction. With the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed. So we can detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein. | ||
+ | |||
+ | ===Characterization=== | ||
+ | Our surface display system can express the Lpp-OmpA-pbrR protein in the outer membrane.So we use the turboid-pbrR to test the pbrR function.With the expression of Lpp-OmpA-pbrR, our bacteria have the ability to adsorb the Pb2+. | ||
+ | [[Image: TP.png|center|frame|100px|<b>Figure 1. TurboID-PbrR</b>]]<br><br> | ||
+ | |||
+ | Our purpose is to detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.And with the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed. | ||
+ | [[Image: TP2.png|center|frame|100px|<b>Figure 2. The different concentration of the Pb2+ in the culture medium </b> (A)0mM;(B)1μM;(C)10μM;(D) 1mM.DAPI channel was used the concentration of the bacteria,the Cy3 channel was used to verify the quantity of the pbrR protein which is not combined with the Pb2+.The result showed that the number of the monomer pbrR protein decreased when there is a higher concentration of the Pb2+.This provided us with the possibility to evaluate the function of our surface display system. ]]<br><br> | ||
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Revision as of 07:32, 12 October 2022
PbrR location detection
This component is made up of TurboID and PbrR. TurboID, a pair of biotin ligases, was designed to overcome the problem of slow existing biotin ligases.PbrR is a lead ion adsorption protein.
Usage and Biology
The PbrR without absorbing lead ions was connected and formed a complex under TurbolD's traction. With the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed. So we can detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.
Characterization
Our surface display system can express the Lpp-OmpA-pbrR protein in the outer membrane.So we use the turboid-pbrR to test the pbrR function.With the expression of Lpp-OmpA-pbrR, our bacteria have the ability to adsorb the Pb2+.
Our purpose is to detect the quantity of the monomer pbrR protein which can combine with the turboid-pbrR fusion protein.And with the combination of the biotin and streptavdin-cy3, the fluorescent signal can be observed.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 1486
Illegal PstI site found at 1428 - 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal SpeI site found at 1486
Illegal PstI site found at 1428 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 1486
Illegal PstI site found at 1428 - 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 1486
Illegal PstI site found at 1428 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1219