Difference between revisions of "Part:BBa K5115038"
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===Introduction===
 
===Introduction===
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This composite part combines [https://parts.igem.org/Part:BBa_K5115035 BBa_K5115035(ribozyme+RBS+MTA+stem-loop)],[https://parts.igem.org/Part:BBa_K5115036 BBa_K5115036(ribozyme+RBS+hpn+stem-loop)]and [https://parts.igem.org/Part:BBa_K5115033 BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop)] in our ribozyme-assisted polycistronic co-expression system：pRAP. To get more information about pRAP, please check [https://2022.igem.wiki/fudan/parts part wiki of 2022 Fudan iGEM ].
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This composite part combines [https://parts.igem.org/Part:BBa_K5115035 BBa_K5115035(ribozyme+RBS+MTA+stem-loop)],[https://parts.igem.org/Part:BBa_K5115036 BBa_K5115036(ribozyme+RBS+hpn+stem-loop)]and [https://parts.igem.org/Part:BBa_K5115033 BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop)] in our ribozyme-assisted polycistronic co-expression system：pRAP. The RNA sequence of ribozyme before the cds can conduct self-cleaving in mRNA, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons in vivo.Self-interaction of the polycistron can be avoid and each cistron can initiate translation with comparable efficiency.<ref>Eiler, D., Wang, J., & Steitz, T. A. (2014). Structural basis for the fast self-cleavage reaction catalyzed by the twister ribozyme. Proceedings of the National Academy of Sciences, 111(36), 13028–13033. https://doi.org/10.1073/pnas.1414571111</ref> To protect the mono-cistron mRNA from degradation, a stem-loop structure is placed at the 3' end of the CDSs.<ref>Liu, Y., Wu, Z., Wu, D., Gao, N., & Lin, J. (2022). Reconstitution of Multi-Protein Complexes through Ribozyme-Assisted Polycistronic Co-Expression. ACS Synthetic Biology, 12(1), 136–143. https://doi.org/10.1021/acssynbio.2c00416</ref>
  
 
MTA is a protein that can bind with nickel ions to reduce its toxity to the 'E.coli'. The hpn is a protein that can  
 
MTA is a protein that can bind with nickel ions to reduce its toxity to the 'E.coli'. The hpn is a protein that can  

Revision as of 18:09, 30 September 2024


ribozyme connected: MTA, Hpn, RcnR_C35L

contributed by Fudan iGEM 2023

Introduction

This composite part combines BBa_K5115035(ribozyme+RBS+MTA+stem-loop),BBa_K5115036(ribozyme+RBS+hpn+stem-loop)and BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop) in our ribozyme-assisted polycistronic co-expression system：pRAP. The RNA sequence of ribozyme before the cds can conduct self-cleaving in mRNA, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons in vivo.Self-interaction of the polycistron can be avoid and each cistron can initiate translation with comparable efficiency.[1] To protect the mono-cistron mRNA from degradation, a stem-loop structure is placed at the 3' end of the CDSs.[2]

MTA is a protein that can bind with nickel ions to reduce its toxity to the 'E.coli'. The hpn is a protein that can sequester metals that accumulate internally to reduce nickel's toxity to the 'E.coli'. RcnR_C35L can regulate the nickel ion channel proteins in the cell membrane to tune the nickel ion transport rate.

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 935
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 198
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. ↑ Eiler, D., Wang, J., & Steitz, T. A. (2014). Structural basis for the fast self-cleavage reaction catalyzed by the twister ribozyme. Proceedings of the National Academy of Sciences, 111(36), 13028–13033. https://doi.org/10.1073/pnas.1414571111
  2. ↑ Liu, Y., Wu, Z., Wu, D., Gao, N., & Lin, J. (2022). Reconstitution of Multi-Protein Complexes through Ribozyme-Assisted Polycistronic Co-Expression. ACS Synthetic Biology, 12(1), 136–143. https://doi.org/10.1021/acssynbio.2c00416