Difference between revisions of "Part:BBa C0062:Experience"

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<partinfo>BBa_R0062 AddReview 4</partinfo>
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<I>ETH Zurich 2014</I>
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= Characterization of two-order crosstalk =
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== Background information ==
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=== System considered ===
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=== Modeling crosstalk ===
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== First-order crosstalk ==
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=== First Level crosstalk: LuxR binds to different HSL and activates the promoter Plux ===
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[[File:ETH Zurich 1crosstalkPlux.png|400px|center]]
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=== Second Level crosstalk: LuxR binds to 3OC6-HSL, its natural HSL, and activates different promoter ===
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[[File:ETH Zurich 2crosstralkRlux.png|400px|center]]
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== Second order crosstalk: Combination of both cross-talk levels  ==
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Other regulatory proteins, like LuxR, bind to different HSL and activates the promoter.
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== Results ==
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Revision as of 17:35, 20 October 2014

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_C0062

User Reviews

UNIQ9b42ea91dc24df8f-partinfo-00000000-QINU


••••

ETH Zurich 2014

Characterization of two-order crosstalk

Background information

System considered

Modeling crosstalk

First-order crosstalk

First Level crosstalk: LuxR binds to different HSL and activates the promoter Plux

ETH Zurich 1crosstalkPlux.png

Second Level crosstalk: LuxR binds to 3OC6-HSL, its natural HSL, and activates different promoter

ETH Zurich 2crosstralkRlux.png

Second order crosstalk: Combination of both cross-talk levels

Other regulatory proteins, like LuxR, bind to different HSL and activates the promoter.

Results


•••••

SUN(Tsinghua)

Part was sequenced and functional. LuxR was used in our Portable Pathogen Detector.

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•••••

wmholtz

Using this part, I have successfully constructed and tested a quorum sensing circuit in E. coli.

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•••••

Youri

This part was used and tested as a subpart in K546000, K546001, K546002, K546003, K546005 and K546546. This part functioned in all cases.

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Kevin (iGEM Braunschweig 2013)

The plasmid pSB1C3 BBa_C0062 from the 2013 distribution Kit was transformed in E. coli XL1 BlueMRF. Sequencing with standard verification primer VF2 confirmed matching sequence of backbone DNA up to the EcoRI restriction site. The rest of the sequence (not shown) does not match the registry entry. 

                   96                                             145
pSB1C3 LuxR   (96) GAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATT
  C0062 VF2    (1) GAGGCAGAATTTCAGATAAAAAAAATCCTTAGCTTTCGCTAAGGATGATT

                   146                                            195
pSB1C3 LuxR  (146) TCTGGAATTCGCGGCCGCTTCTAGAGATGAAAAACATAAATGCCGACGAC
  C0062 VF2   (51) TCTGGAATTCGACGCAA-TGGGTGCGCTGTCTACTAAATACAACGACACC

                   196                                            245
pSB1C3 LuxR  (196) ACATACAGAATAATTAATAAAATTAAAGCTTGTAGAAGCAATAATGATAT
  C0062 VF2  (100) CCGGAAAAAGCCTCCCGTACTTACGACGCTCACCGTGACGGTTTCGTTAT

                   246
pSB1C3 LuxR  (246) TAATCAATGC...
  C0062 VF2  (150) CGCTGGCGGC...


A restriction assay (Figure 1) showed that the sequenced part has no XbaI restriction site following the EcoRI site indicating another part in front of BBa_C0062 with a length of at least 1000 bp.

Figure 1: restriction assay of BBa_C0062 with the indicated restriction enzymes

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UNIQ9b42ea91dc24df8f-partinfo-00000007-QINU