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)], 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*.
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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)] . 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] 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''.
  
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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.
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With this design, we achieve co-expression of [https://parts.igem.org/Part:BBa_K5115050 MTA], [https://parts.igem.org/Part:BBa_K1151001 hpn], [https://parts.igem.org/Part:BBa_K5115000 RcnR C35L] 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===
 
===Characterization===

Revision as of 00:07, 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. By inserting ribozyme sequences 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, hpn, RcnR C35L 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