Part:BBa_K5115042
ribozyme connected: MTA, RcnR_C35L, Hpn
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 642
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 198
- 1000COMPATIBLE WITH RFC[1000]
References
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