Difference between revisions of "Part:BBa K1442023"
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<p> This is an Internal Ribosome Entry Site (IRES) acting in mammalian cells. It allows translation initiation in the middle of an RNA strand which is required for the fuctioning of our replicon as there are multiple protein coding regions separated by structural nucleotide sequences interspersed throuhgout the RNA strand. We chose to investigate two IRES sequences to compare the relative efficiency and determine which was better for incorporation into our system. These were the Encephalomyocarditis virus (EMCV) and the IRES derived from the NF-kappaB repressor factor (NKRF) untranslated region.</p> | <p> This is an Internal Ribosome Entry Site (IRES) acting in mammalian cells. It allows translation initiation in the middle of an RNA strand which is required for the fuctioning of our replicon as there are multiple protein coding regions separated by structural nucleotide sequences interspersed throuhgout the RNA strand. We chose to investigate two IRES sequences to compare the relative efficiency and determine which was better for incorporation into our system. These were the Encephalomyocarditis virus (EMCV) and the IRES derived from the NF-kappaB repressor factor (NKRF) untranslated region.</p> | ||
<p> IRES' were first discovered in EMCV and polioviruses and the EMCV has been the favoured IRES for use in replicon systems such as Lohmann (1999) and Lee (2005).</p> | <p> IRES' were first discovered in EMCV and polioviruses and the EMCV has been the favoured IRES for use in replicon systems such as Lohmann (1999) and Lee (2005).</p> | ||
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<p> IRESes are commonly used by viruses to ensure translation is active even during times when host translation is inhibited. This is ideal for our system as in order to maintain the RNA in the cell for the maximum length of time the translation of the RdRp should be constant to balance the instability and subsequent degradation of the RNA strands. </p> | <p> IRESes are commonly used by viruses to ensure translation is active even during times when host translation is inhibited. This is ideal for our system as in order to maintain the RNA in the cell for the maximum length of time the translation of the RdRp should be constant to balance the instability and subsequent degradation of the RNA strands. </p> | ||
<p> The EMCV IRES functions via its secondary structure which attracts ribosomes to the vicinity of the start codon of the protein coding region of the RNA. | <p> The EMCV IRES functions via its secondary structure which attracts ribosomes to the vicinity of the start codon of the protein coding region of the RNA. | ||
The EMCV IRES forms a complex secondary structure with many hairpin loops and | The EMCV IRES forms a complex secondary structure with many hairpin loops and | ||
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| + | ===Usage and Biology=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 14:27, 6 October 2014
EMCV IRES
This is an Internal Ribosome Entry Site (IRES) acting in mammalian cells. It allows translation initiation in the middle of an RNA strand which is required for the fuctioning of our replicon as there are multiple protein coding regions separated by structural nucleotide sequences interspersed throuhgout the RNA strand. We chose to investigate two IRES sequences to compare the relative efficiency and determine which was better for incorporation into our system. These were the Encephalomyocarditis virus (EMCV) and the IRES derived from the NF-kappaB repressor factor (NKRF) untranslated region.
IRES' were first discovered in EMCV and polioviruses and the EMCV has been the favoured IRES for use in replicon systems such as Lohmann (1999) and Lee (2005).
IRESes are commonly used by viruses to ensure translation is active even during times when host translation is inhibited. This is ideal for our system as in order to maintain the RNA in the cell for the maximum length of time the translation of the RdRp should be constant to balance the instability and subsequent degradation of the RNA strands.
The EMCV IRES functions via its secondary structure which attracts ribosomes to the vicinity of the start codon of the protein coding region of the RNA. The EMCV IRES forms a complex secondary structure with many hairpin loops and Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]