Difference between revisions of "Part:BBa K5115035"
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===Introduction===
 
===Introduction===
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This composite part is composed of MTA and improved ribozyme-assisted polycistronic co-expression system: pRAP. The MTA is a protein that can bind with nickel ion and reduce its toxity, get details about MTA in  [https://parts.igem.org/Part:BBa_K5115050 BBa_K5115050(MTA, Metallothionein)].  
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This composite part is composed of MTA and improved ribozyme-assisted polycistronic co-expression system：pRAP. We introduced this ribozyme-assisted polycistronic co-expression system in [https://2022.igem.wiki/fudan/parts 2022]. By inserting [https://parts.igem.org/Part:BBa_K4765020 ribozyme sequences] before MTA, the RNA sequences of Twister ribozyme conduct self-cleaving in the mRNA..<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.</ref> To protect the mono-cistron mRNA from degradation, a stem-loop structure is placed at the 3' end of MTA.<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.</ref> The Metallothioneins (MTA) are intracellular, low molecular, low molecular weight, cysteine-rich proteins. Ubiquitous in eukaryotes, MTA have unique structural characteristics to give potent metal-binding and redox capabilities.<ref>Coyle, P., Philcox, J. C., Carey, L. C., & Rofe, A. M. (2002). Metallothionein: The multipurpose protein. Cellular and Molecular Life Sciences: CMLS, 59(4), 627–647.</ref>
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===Usage and Biology===
 
===Usage and Biology===
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The ribozyme-assisted polycistronic co-expression system can ensure that each cistron can initiate translation with comparable efficiency. For more information, please check [https://2022.igem.wiki/fudan/parts part wiki of 2022 Fudan iGEM].
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The heterologously expressed codon-optimized MTA can endowing ''E.coli'' with detoxifying capability.
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Get details in [https://parts.igem.org/Part:BBa_K5115050 BBa_K5115050]
  
  

Revision as of 00:28, 1 October 2024


ribozyme+RBS+MTA+stem-loop

contributed by Fudan iGEM 2023

Introduction

This composite part is composed of MTA and improved ribozyme-assisted polycistronic co-expression system：pRAP. We introduced this ribozyme-assisted polycistronic co-expression system in 2022. By inserting ribozyme sequences before MTA, the RNA sequences of Twister ribozyme conduct self-cleaving in the mRNA..[1] To protect the mono-cistron mRNA from degradation, a stem-loop structure is placed at the 3' end of MTA.[2] The Metallothioneins (MTA) are intracellular, low molecular, low molecular weight, cysteine-rich proteins. Ubiquitous in eukaryotes, MTA have unique structural characteristics to give potent metal-binding and redox capabilities.[3]


Usage and Biology

The heterologously expressed codon-optimized MTA can endowing E.coli with detoxifying capability.

Get details in BBa_K5115050


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 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.
  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.
  3. ↑ Coyle, P., Philcox, J. C., Carey, L. C., & Rofe, A. M. (2002). Metallothionein: The multipurpose protein. Cellular and Molecular Life Sciences: CMLS, 59(4), 627–647.