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Revision as of 16:11, 16 October 2018

COMPOSITE PART


Abstract Thermosensor sfGFP

Composite Part

  • Abstract

    This year, we added a total of 7 composite parts, including heat-inducible RNA-based thermosensor measurement device, heat-repressible RNA-based thermosensor measurement device, cold-inducible RNA-based thermosensor measurement device, cold-repressible RNA-based thermosensors measurement device, sfGFP_optimism measurement device, superfolder GFP measurement device and sfGFP(BbsI free)measurement device.

  • Thermosensor measurement device

    Part Name Part Number
    Heat-inducible RNA-based thermosensor measurement device BBa_K2541405
    Heat-repressible RNA-based thermosensor measurement device BBa_K2541406
    Cold-repressible RNA-based thermosensor measurement device BBa_K2541407
    Cold-inducible RNA-based thermosensor measurement device BBa_K2541408

    The thermosensor sequence is constructed on the pSB1C3 vector by Golden Gate assembly. The measurement device is composed of Anderson promotor (BBa_J23104), thermosensor (BBa_K2541051), sfGFP_optimism (BBa_K2541400) and double terminator (BBa_B0010 and BBa_B0012). We select a constitutive Anderson promoter J23104 as an appropriate promoter by pre-experiment. The sfGFP_optimism has faster folding speed and higher fluorescence intensity. The double terminator can reduce leakage (Figure 1). We characterized RNA-based thermosensors in E.coli DH5a.

    Figure 3. Diagram of the measurement device.

  • sfGFP Measurement Device

    In addition, we combined our cold-induced RNA thermosensors (cspA mRNA) with the promoter J23104 as a composite part for uploading, which is more conducive to other users directly, users can also only use a cold-induced RNA thermosensor according to their own needs.

    Part Name Part Number
    sfGFP_optimism measurement device BBa_K2541402
    superfolder GFP measurement device BBa_K2541403
    sfGFP(BbsI free) measurement device BBa_K2541404

    Introduction

    They are three measurement device for sfGFP_optimism (BBa_K2541400), superfolder GFP (BBa_I746916) and sfGFP(BbsI free)(BBa_K2541401) fluorescence intensity, which is composed of Anderson promoter (BBa_J23104), RBS (BBa_B0034), sfGFP and double terminator (BBa_B0010 and BBa_B0012).