Team:SUSTech Shenzhen/Contribution

  • SUSTech_Shenzhen

    • Contribution

    Parts we used and improved and contribution to the field development.

    Contribution

    Contribution

    We have used the BBa_B0014 terminator Acella in our gene design(figure. 1). It’s a double terminator which would terminate the transcription in both directions. There are some potential advantages for a double terminator. Firstly, a double terminator may consume fewer resources because it can terminate two gene expressions at the same time. Secondly, it could spontaneously prevent the expression because of the reverse arrangement of genes.

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    Figure. 1 PhlF + GFP + PphlF sequence design.

    We successfully verified the availability of this bidirectional terminator in E. coli Acella. In our experiment, both PhlF and GFP were expressed with the assistance of this terminator in E. coli Acella system (figure.2, figure.3). And the correct size of PhlF from western blot indicates an effective termination (figure.3).

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    Figure. 2 Western blot of PhlF (E is experiment; C is control; S is supernatant; P is precipitate; PC is positive control).

    The above contributions may provide some validation and suggestions for the following teams who want to use double terminators for their circuit design.

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    Figure. 3 Fluorescence test of GFP with different concentrations of IPTG and same concentration of AHL.

    Parts

    Part Number type Basic/composite length
    transcription terminator T1 from the E. coli rrnB gene BBa_K3914001 Terminator Basic 72
    phage T7 early transcription terminator BBa_K3914002 Terminator Basic 28
    Regulator of capsule synthesis B BBa_K3914003 Coding basic 669
    transcription terminator from phage lambda BBa_K3914004 Terminator basic 95
    lac operon BBa_K3914005 lac operon basic 1493
    Esa box of EsaR protein BBa_K3914006 Reporter basic 18
    Ribosome bingding site of Esa box BBa_K3914007 RBS basic 12
    Green fluorescence protein GFP BBa_K3914008 Coding basic 714
    PhlF Repressible Promoter PphlF BBa_K3914009 Regulatory basic 45
    Transcriptional repressor PhlF BBa_K3914010 Coding basic 621
    the detector protein of vibrio cholerae quorum sensing signal CAI-1 BBa_K3914013 Coding basic 2058
    a downstream phospholated protein of LuxU, and the regulator of tpQrr4 BBa_K3914014 Coding basic 1392
    a downstream phospholated protein of CqsS, and the upstream protein of LuxO BBa_K3914015 Coding basic 366
    the selective promotor induced by LuxO BBa_K3914016 Coding basic 31
    RNAseH II in Lactococcus lactis BBa_K3914034 Coding basic 556
    rrnb T1 and rrnb T2 double terminator BBa_K3914035 Terminator basic 326
    N-acetylmuramidase gene AcmA BBa_K3914036 Coding basic 1376
    open reading frame BBa_K3914037 Coding basic 285
    gadR Cl−-dependent positive regulator with rrnBT1T2 terminator BBa_K3914038 Regulatory basic 1367
    AHLs synthesis gene luxI BBa_K3914039 Coding basic 633
    Erythromycin Constitutive Promoter BBa_K3914040 Regulatory basic 536
    lac operated PhlF synthesis circuit BBa_K3914101 Regulatory composite 2270
    PphlF mediated GFP testing antibacterial circuit BBa_K3914102 Coding composite 907
    Phlf mediated GFP testing antibacterial circuit BBa_K3914103 Coding composite 3177
    plasmid lac operon induced RcsB BBa_K3914104 Composite composite 2399
    the CAI-1 detector engineering sequences BBa_K3914105 Composite composite 4421
    the designing plasmid to detect CAI-1 BBa_K3914109 Plasmid composite 78
    L.reuteri AHL synthesize pathway BBa_K3914113 Composite composite 3320
    L.reuteri autophagy pathway BBa_K3914114 Composite composite 1175
    pBbE6k plasmid backbone BBa_K3914128 Plasmid_Backbone composite 78
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