Difference between revisions of "Part:BBa K5115042"
 
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<partinfo>BBa_K5115042 short</partinfo>
 
<partinfo>BBa_K5115042 short</partinfo>
  
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<html><img style="float:right;width:128px" src="https://static.igem.wiki/teams/5115/czh/mineral-logo.svg" alt="contributed by Fudan iGEM 2023"></html>
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<html><img style="float:right;width:128px" src="https://static.igem.wiki/teams/5115/czh/mineral-logo.svg" alt="contributed by Fudan iGEM 2024"></html>
 
__TOC__
 
__TOC__
 
===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_K5115033 BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop)] and [https://parts.igem.org/Part:BBa_K5115036 BBa_K5115036(ribozyme+RBS+hpn+stem-loop)]. We introduced this ribozyme-assisted polycistronic co-expression system from [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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This composite part combines [https://parts.igem.org/Part:BBa_K5115035 BBa_K5115035(ribozyme+RBS+MTA+stem-loop)], [https://parts.igem.org/Part:BBa_K5115033 BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop)] and [https://parts.igem.org/Part:BBa_K5115036 BBa_K5115036(ribozyme+RBS+Hpn+stem-loop)]. We introduced this ribozyme-assisted polycistronic co-expression system from [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 [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 toxicity to the ''E.coli''. The hpn is a protein that can sequester metals that accumulate internally to reduce nickel's toxicity 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 toxicity to the ''E.coli''. The Hpn is a protein that can sequester metals that accumulate internally to reduce nickel's toxicity to the ''E.coli''. RcnR C35L can regulate the nickel ion channel proteins in the cell membrane to tune the nickel ion transport rate.
  
 
===Usage and Biology===
 
===Usage and Biology===
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To ascertain which order the three proteins should be assembled for the best results, we create six different parts containing all the sequential possibilities. We finally confirm that [https://parts.igem.org/Part:BBa_K5115038 ribozyme connected: MTA, hpn, RcnR_C35L] works the best, please check it for experiment results.
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To ascertain which order the three proteins should be assembled for the best results, we create six different parts containing all the sequential possibilities. We finally confirm that the order of the three parts don't affect much on their effect. We choose to use [https://parts.igem.org/Part:BBa_K5115038 ribozyme connected: MTA, Hpn, RcnR_C35L] at last, please check it for the experiment results.
  
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===Sequence and Features===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Latest revision as of 12:12, 2 October 2024


ribozyme connected: MTA, RcnR_C35L, Hpn

contributed by Fudan iGEM 2024

Introduction

This composite part combines BBa_K5115035(ribozyme+RBS+MTA+stem-loop), BBa_K5115033(ribozyme+RBS+RcnR_C35L+stem-loop) and BBa_K5115036(ribozyme+RBS+Hpn+stem-loop). We introduced this ribozyme-assisted polycistronic co-expression system from 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 toxicity to the E.coli. The Hpn is a protein that can sequester metals that accumulate internally to reduce nickel's toxicity to the E.coli. RcnR C35L can regulate the nickel ion channel proteins in the cell membrane to tune the nickel ion transport rate.

Usage and Biology

To ascertain which order the three proteins should be assembled for the best results, we create six different parts containing all the sequential possibilities. We finally confirm that the order of the three parts don't affect much on their effect. We choose to use ribozyme connected: MTA, Hpn, RcnR_C35L at last, please check it for the experiment results.

Sequence and Features

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 642
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 198
  • 1000
    COMPATIBLE WITH RFC[1000]


References