Team:Jilin China/Basic Part

BASIC PART


Basic Part

  • Abstract

    This year, Jilin_China added 91 basic parts to the registry, including heat-inducible RNA thermosensors, heat-repressible RNA thermosensors, cold-inducible RNA thermosensors, cold-repressible RNA thermosensors and two different sfGFP. We have characterized and measured all of these parts, calculated their melting temperatures by using mathematical modeling, and built a synRT toolkit that allows users to select the appropriate RNA thermosensors in artificial biological systems.

    Our team's favorite basic part is the codon-optimized sfGFP (BBa_K2541400) and will be introduced in detail below:

  • sfGFP_optimism (BBa_K2541400)

    sfGFP (superfolder GFP), which emission and excitation wavelength are similar to GFP, but its fluorescence intensity and folding speed are higher than GFP. Because of its advantages, our measurement device used sfGFP as the reporter protein. However, since we use Goldengate assembly this year, the existing sfGFP (BBa_I746916) in the registry contains a BbsI endonuclease cutting site. We designed a site-directed mutation of sfGFP (BBa_K2541401), made a double-base mutation to the BbsI recognition site without changing its amino acid sequence. BBa_K2541401 is completely suitable for Goldengate assembly, so we also call it sfGFP for Goldengate.

    There's still one problem that we cannot predict the effect of the sfGFP after mutation. In addition, we found a codon optimized sfGFP for prokaryote, and called it sfGFP_optimism. Then we designed a composite part, which contains J23103, B0034 and sfGFP_optimism, and did experiment to compare it with sfGFP and sfGFP for goldengate. As the result shows, the sfGFP_optimism has higher fluorescence intensity than others, so we finally chose sfGFP_optimism as our reporter gene.

    You can see the experiment results in the improvement page. Click Here!

    Since sfGFP has more advantages than GFP, and Goldengate assembly will be used by more researchers as an efficient and scarless assembly method in the future, so we decided to add the sfGFP_optimism to the parts registry.

  • RNA thermosensors

    Heat-inducible RNA thermosensors

    Heat-repressible RNA thermosensors

    Cold-inducible RNA thermosensors

    Cold-repressible RNA thermosensors

  • Basic parts content