Difference between revisions of "Part:BBa K4204013"
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===Description=== | ===Description=== | ||
| − | The protein quality control system (Pro-QC system) is universal that can be applied to all kinds of protein expression. The system contains a toehold switch (BBa_K4204005) at the 5' end of an mRNA and a corresponding trigger (Cis-trigger) (BBa_K4204006)sequence with reverse complementarity to the toehold switch at the 3' end of the mRNA. The switch binds with its complementary sequence and blocks the RBS inside a neck-ring structure; this blocks transcription initiation. When Cis-trigger appears, the switch will bind with the trigger preferentially because the Cis-trigger has a higher affinity with the switch. Thus, the switch and the trigger RNA pairs deformed the neck-ring structure to free the RBS and circularized the mRNA regardless of the gene of interest The exposed RBS enables translation initiation. Thus only the full-length mRNAs with trigger sequence at its 3’end are selectively used as the templates for translation. | + | The protein quality control system (Pro-QC system) is universal that can be applied to all kinds of protein expression. The system contains a toehold switch (BBa_K4204005) at the 5' end of an mRNA and a corresponding trigger (Cis-trigger) (BBa_K4204006)sequence with reverse complementarity to the toehold switch at the 3' end of the mRNA. The switch binds with its complementary sequence and blocks the RBS inside a neck-ring structure; this blocks transcription initiation. When Cis-trigger appears, the switch will bind with the trigger preferentially because the Cis-trigger has a higher affinity with the switch. Thus, the switch and the trigger RNA pairs deformed the neck-ring structure to free the RBS and circularized the mRNA regardless of the gene of interest The exposed RBS enables translation initiation. Thus only the full-length mRNAs with trigger sequence at its 3’end are selectively used as the templates for translation. Since Lac operon is the most common induction system used for protein expression, our team improved upon it and uploaded the Protein quality control system with GFP between the switch and trigger on pET-28a(+) (a common expression vector with His-tag and Lac operator) to the part registry (in which the GFP could be replaced by other parts). Thus, other IGEM teams can use the protein quality control system. |
===Sequence and Features=== | ===Sequence and Features=== | ||
<partinfo>BBa_K4204013 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4204013 SequenceAndFeatures</partinfo> | ||
Revision as of 14:16, 6 October 2022
iPTG+sfGFP+quality control system
Description
The protein quality control system (Pro-QC system) is universal that can be applied to all kinds of protein expression. The system contains a toehold switch (BBa_K4204005) at the 5' end of an mRNA and a corresponding trigger (Cis-trigger) (BBa_K4204006)sequence with reverse complementarity to the toehold switch at the 3' end of the mRNA. The switch binds with its complementary sequence and blocks the RBS inside a neck-ring structure; this blocks transcription initiation. When Cis-trigger appears, the switch will bind with the trigger preferentially because the Cis-trigger has a higher affinity with the switch. Thus, the switch and the trigger RNA pairs deformed the neck-ring structure to free the RBS and circularized the mRNA regardless of the gene of interest The exposed RBS enables translation initiation. Thus only the full-length mRNAs with trigger sequence at its 3’end are selectively used as the templates for translation. Since Lac operon is the most common induction system used for protein expression, our team improved upon it and uploaded the Protein quality control system with GFP between the switch and trigger on pET-28a(+) (a common expression vector with His-tag and Lac operator) to the part registry (in which the GFP could be replaced by other parts). Thus, other IGEM teams can use the protein quality control system.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 64
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 759
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 811
- 1000COMPATIBLE WITH RFC[1000]