Difference between revisions of "Part:BBa K5398002"
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<p> <b>The mechanism of cmRNA </b> </p>
 
<p> <b>The mechanism of cmRNA </b> </p>
 
<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>
 
<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> iGEM Gifu 2014 also used a similar part (BBa_K1332005). If you want to learn more about it, please click the link above. https://parts.igem.org/Part:BBa_K1332005 </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>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 04:02, 7 September 2024


The 5' intron of td gene from T4 phage

This part is a part of the circular mRNA (cmRNA) (3´ side).

The mechanism of cmRNA

To generate the squid ring proteins with various long tandem repeats (TRn), we utilized the td intron, an intron of the td 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.

iGEM Gifu 2014 also used a similar part (BBa_K1332005). If you want to learn more about the td intron, please click the link above. https://parts.igem.org/Part:BBa_K1332005

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 35
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]