Difference between revisions of "Part:BBa K5398003"
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<partinfo>BBa_K5398003 short</partinfo>
 
<partinfo>BBa_K5398003 short</partinfo>
  
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<p>This part is a part of the circular mRNA (cmRNA) (5' side). </p>
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<p>This part is a component of the <i>td</i> intron, an intron of the <i>td</i> gene from T4 phage belonging to group IE, which can form a circular mRNA (cmRNA) to make the ribosomes repeatedly translate the extron. This strategy can be used for expression of long peptides containing tandem repeats, like fibrous proteins.</p>
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<p> <b>The mechanism of cmRNA </b> </p>
 
<p> <b>The mechanism of cmRNA </b> </p>
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<p>To generate the squid ring proteins with various long tandem repeats (TRn), we utilized the <i>td</i> intron, an intron of the <i>td</i> gene from T4 phage belonging to group IE, which circularizes the exon to form a back-splice junction (BSJ) in a reaction catalyzed by guanosine. To ensure that the ribosomes do not translate the ORF of TRn from unprocessed linear mRNA, the ribosome binding sequence (RBS) and translation start codon ATG were placed downstream of the TRn coding sequence. Consequently, the regulatory sequences were located upstream of the coding sequence only after circularization of the mRNA. To purify the resulting TRn polypeptides, a His tag was incorporated into the ORF. If the mRNA is circularized, the ribosome could circle the cmRNA, producing a long repeating polypeptide. </p>
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<P>Due to the special internal structure, the <i>td</i> intron can circularize the exon to form a back-splice junction (BSJ) in a reaction catalyzed by guanosine. To ensure that the ribosomes do not translate the ORF of gene of interest (GOI) from unprocessed linear mRNA, the ribosome binding sequence (RBS) and start codon ATG were placed downstream of GOI coding sequence. Consequently, the regulatory sequences were located upstream of the coding sequence only after circularization of the mRNA. To purify the resulting polypeptides, a His tag was incorporated into the GOI. If the mRNA is circularized, the ribosome could circle the cmRNA, producing a long repeating polypeptide (Fig. 2). </p>
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<p> iGEM Gifu 2014 also used a similar part (BBa_K1332005). If you want to learn more about the <i>td</i> intron, please click the link above. https://parts.igem.org/Part:BBa_K1332005 </p>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 09:18, 10 September 2024


The 3' intron of td gene from T4 phage

This part is a component of the td intron, an intron of the td gene from T4 phage belonging to group IE, which can form a circular mRNA (cmRNA) to make the ribosomes repeatedly translate the extron. This strategy can be used for expression of long peptides containing tandem repeats, like fibrous proteins.

The mechanism of cmRNA

Due to the special internal structure, the td intron can circularize the exon to form a back-splice junction (BSJ) in a reaction catalyzed by guanosine. To ensure that the ribosomes do not translate the ORF of gene of interest (GOI) from unprocessed linear mRNA, the ribosome binding sequence (RBS) and start codon ATG were placed downstream of GOI coding sequence. Consequently, the regulatory sequences were located upstream of the coding sequence only after circularization of the mRNA. To purify the resulting polypeptides, a His tag was incorporated into the GOI. If the mRNA is circularized, the ribosome could circle the cmRNA, producing a long repeating polypeptide (Fig. 2).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 217
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]