Difference between revisions of "Part:BBa K5115038"
Line 6: Line 6:
 
__TOC__
 
__TOC__
 
===Introduction===
 
===Introduction===
−
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>
+
This composite part combines [https://parts.igem.org/Part:BBa_K5115035 BBa_K5115035(ribozyme+RBS+MTA+stem-loop)], BBa_K5115036(ribozyme+RBS+hpn+stem-loop)and BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop) . We introduced this ribozyme-assisted polycistronic co-expression system in [2022](https://2022.igem.wiki/fudan/parts). By inserting [ribozyme sequences](https://parts.igem.org/Part:BBa_K4765020) between CDSs in a polycistron, the RNA sequences of Twister ribozyme conduct self-cleaving, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons *in vivo*.
  
−
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  
+
With this design, we achieve co-expression of [MTA](https://parts.igem.org/Part:BBa_K5115050), Hpn 没有它的basic, [RcnR C35L](https://parts.igem.org/Part:BBa_K5115000) at similar level. 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.
−
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===
 
===Characterization===

Revision as of 00:02, 1 October 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) . We introduced this ribozyme-assisted polycistronic co-expression system in [2022](https://2022.igem.wiki/fudan/parts). By inserting [ribozyme sequences](https://parts.igem.org/Part:BBa_K4765020) between CDSs in a polycistron, the RNA sequences of Twister ribozyme conduct self-cleaving, and the polycistronic mRNA transcript is thus co-transcriptionally converted into individual mono-cistrons *in vivo*.

With this design, we achieve co-expression of [MTA](https://parts.igem.org/Part:BBa_K5115050), Hpn 没有它的basic, [RcnR C35L](https://parts.igem.org/Part:BBa_K5115000) at similar level. 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