Difference between revisions of "Part:BBa C0062"

Line 38: Line 38:
 
Author: Adrián Requena Gutiérrez, Carolina Ropero
 
Author: Adrián Requena Gutiérrez, Carolina Ropero
 
<br>
 
<br>
Summary: We adapted the part to be able to assemble transcriptional units with the Golden Gate assembly method
+
Summary:  
 +
We adapted the part to be able to assemble transcriptional units with the Golden Gate assembly method
 
<br>
 
<br>
 
Documentation:
 
Documentation:
 
In order to create our complete [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Printeira Part Collection] of parts compatible with the Golden Gate assembly method, we have made the part [https://parts.igem.org/Part:BBa_K2656016 BBa_K2656016] which is BBa_C0062 adapted to the Golden Gate Technology.
 
In order to create our complete [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Printeira Part Collection] of parts compatible with the Golden Gate assembly method, we have made the part [https://parts.igem.org/Part:BBa_K2656016 BBa_K2656016] which is BBa_C0062 adapted to the Golden Gate Technology.

Revision as of 23:59, 17 October 2018

luxR repressor/activator, (no LVA?)

In complex with HSL, LuxR binds to the Lux promoter, activating transcription from Pr BBa_R0062, and repressing transcription from Pl BBa_R0063.

The lux cassette of V. fischeri contains a left and a right promoter. The right promoter gives weak constitutive expression of downstream genes.This expression is up-regulated by the action of the Lux activator, LuxR complexed to HSL. Two molecules of LuxR protein form a complex with two molecules the signalling compound homoserine lactone (HSL). This complex binds to a palindromic site on the promoter, increasing the rate of transcription.


Usage and Biology

We improved this part by using it in a part which allows for the constitutive expression of LuxR (BBa_K1893004). This part also includes the pLux promoter, so that genes placed downstream the promoter will be expressed when induced with 3OC6-AHL. We also made a part which allows for the characterisation of LuxR, as we placed GFP downstream the pLux promoter (BBa_K1893005). The part is currently being characterised.

Unspecified.

Sequence and Features


Barcodes are discontinued, but one was appended to the sequence of this part. Composite parts using this part will include the barcode. More ...

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]


  • Allergen characterization of BBa_C0062: Not a potential allergen

The Baltimore Biocrew 2017 team discovered that proteins generated through biobrick parts can be evaluated for allergenicity. This information is important to the people using these parts in the lab, as well as when considering using the protein for mass production, or using in the environment. The allergenicity test permits a comparison between the sequences of the biobrick parts and the identified allergen proteins enlisted in a data base.The higher the similarity between the biobricks and the proteins, the more likely the biobrick is allergenic cross-reactive. In the full-length alignments by FASTA, 30% or more amount of similarity signifies that the biobrick has a Precaution Status meaning there is a potential risk with using the part. A 50% or more amount of identity signifies that the biobrick has a Possible Allergen Status. In the sliding window of 80 amino acid segments, greater than 35% signifies similarity to allergens. The percentage of similarity implies the potential of harm biobricks’ potential negative impact to exposed populations. For more information on how to assess your own biobrick part please see the “Allergenicity Testing Protocol” in the following page http://2017.igem.org/Team:Baltimore_Bio-Crew/Experiments


For the biobrick Part BBa_C0062, there was a 0% of identity match and 0% similarity match to the top allergens in the allergen database. This means that the biobrick part is not of potential allergen status. In 80 amino acid alignments by FASTA window, no matches found that are greater than 35% for this biobrick. This also means that there is not of potential allergen status.

Contribution: SiCAU-China 2017

Authors:Wenwen Lin

Summary:The QS system LuxR/LuxI has been widely used in kinds of logic circuit. But this year we find that LuxR can not always work well in E.coli. The LuxR, in E.coli, will not work when E.coli enter into stationary phase. This characteristic may be useful to users who will use LuxR as an activating transcription. (more details)

BBa C0062 SiCAU-China contribution.png

Contribution

Group: Valencia_UPV iGEM 2018
Author: Adrián Requena Gutiérrez, Carolina Ropero
Summary: We adapted the part to be able to assemble transcriptional units with the Golden Gate assembly method
Documentation:

In order to create our complete [http://2018.igem.org/Team:Valencia_UPV/Part_Collection Printeira Part Collection] of parts compatible with the Golden Gate assembly method, we have made the part BBa_K2656016 which is BBa_C0062 adapted to the Golden Gate Technology.