Difference between revisions of "Part:BBa K2462001"
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===Verification===
 
===Verification===
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For the verification of this RBS, we use this RBS to construct a circuit with our dehalogenase RdANP. Due to the limitation of time and experimental equipment, we didn’t do the SDS-PAGE, instead, we did HPLC to test the our eyzeme. We insist the idea that if we succeed in the expression of our enzyme, we shall verify the RBS at the same time. We use the HPLC to test the effects of our enzyme, but we didn’t get convincing results because there are proteins interrupting the estimation.
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For the verification of this RBS, we use this RBS to construct a circuit with our dehalogenase RdANP. Due to the limitation of time and experimental equipment, we didn’t do the SDS-PAGE, instead, we did HPLC to test the our enzyme. We insist the idea that if we succeed in the expression of our enzyme, we shall verify the RBS at the same time. We use the HPLC to test the effects of our enzyme, but we didn’t get convincing results because there are proteins interrupting the estimation.
  
 
The details related to the experiment are described in the part page of BBa K2462000:
 
The details related to the experiment are described in the part page of BBa K2462000:

Revision as of 15:02, 1 November 2017


RBS

Description

This part is a RBS used in Bacillus megaterium. We got its sequence in pSPAsp-hp, which is a plasmid used for protein expression in Bacillus megaterium. Then the whole part has been synthesized by AuGCT.

Verification

For the verification of this RBS, we use this RBS to construct a circuit with our dehalogenase RdANP. Due to the limitation of time and experimental equipment, we didn’t do the SDS-PAGE, instead, we did HPLC to test the our enzyme. We insist the idea that if we succeed in the expression of our enzyme, we shall verify the RBS at the same time. We use the HPLC to test the effects of our enzyme, but we didn’t get convincing results because there are proteins interrupting the estimation.

The details related to the experiment are described in the part page of BBa K2462000:

https://parts.igem.org/Part:BBa_K2462000 In order to make the experiment more convincing, we also made another group of experimrnts. We took bacteria with RdhANP induced by xylose as the experimental group, and noninduced bacteria as the control. Dots colored differently indicate result under different conditions: green for transformed and induced group; blue for untransformed but induced group; yellow for transformed but induced group. The results are depicted as following:


WHU-China-Part-01.png
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]

Disscusion

We decided to take this promoter as our part not only because we used it in our own project but we do hope that the following teams can have more options when they choose to use Bacillus megaterium as their host. We failed to verify this promoter by ourselves, but we are pleasant to give other teams the information of the original source for this promoter. As we get this promoter from Addgene, information can be found in their web. And few articles using this promoter can be found in their page for other teams’ reference.

More details about pSPAsp-hp:

http://www.addgene.org/48122/