Difference between revisions of "Team:Gifu/project/"
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− | <p> The group I intron in td gene of T4 phage has self-splicing mechanism. The self-splicing is a mechanism that circularizes the intron and connects exons. This is catalyzed by several base sequences of the ends of the introns as a ribozyme. We permuted exons and introns with the mechanism and attempted an exon circularization. So we constructed mRNA circularization devices in last year.</p> | + | <p> The group I intron in td gene of T4 phage has self-splicing mechanism. The self-splicing is a mechanism that circularizes the intron and connects exons. This is catalyzed by several base sequences of the ends of the introns as a ribozyme. We permuted exons and introns with the mechanism and attempted an exon circularization. So we constructed mRNA circularization devices in last year.</p> |
<h2>The circularization mechanism of group I intron</h2> | <h2>The circularization mechanism of group I intron</h2> | ||
− | <p>The group I intron is capable of self-splicing. The mRNA circularization device is based on the mechanism. We explain the circularization mechanism of group I intron with td gene of T4 phage as an example. Td gene consists of an upstream exon, an upstream intron, an ORF, a downstream intron and a downstream exon. | + | <p> The group I intron is capable of self-splicing. The mRNA circularization device is based on the mechanism. We explain the circularization mechanism of group I intron with td gene of T4 phage as an example. Td gene consists of an upstream exon, an upstream intron, an ORF, a downstream intron and a downstream exon. |
As the first step, a nucleophilic attack by a guanosine separates the upstream exon from the upstream intron and then the guanosine bonds to the 5’ end of the upstream intron. | As the first step, a nucleophilic attack by a guanosine separates the upstream exon from the upstream intron and then the guanosine bonds to the 5’ end of the upstream intron. | ||
− | As the second step, the downstream exon is separated from the downstream intron by a nucleophilic attack. The nucleophilic attack takes place by a hydroxy group at the 3’ end of the upstream exon. (Figure 1)<br> | + | As the second step, the downstream exon is separated from the downstream intron by a nucleophilic attack. The nucleophilic attack takes place by a hydroxy group at the 3’ end of the upstream exon. (Figure 1)<br> |
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<img src="https://static.igem.org/mediawiki/2014/4/41/SS1_GIFU.png" width="700px"></img><br> | <img src="https://static.igem.org/mediawiki/2014/4/41/SS1_GIFU.png" width="700px"></img><br> | ||
<b>Fig.2 Self-splicing in T4 phage: the first and second step (Blue: intron, Orange: exon)</b><br><br> | <b>Fig.2 Self-splicing in T4 phage: the first and second step (Blue: intron, Orange: exon)</b><br><br> | ||
− | As the third step, the upstream intron bonds to the downstream intron by an attack on an adenine of the upstream intron. The attack takes place by a hydroxyl group of an end of the downstream intron. And then a circular intron is formed.(Figure 2)<br> | + | As the third step, the upstream intron bonds to the downstream intron by an attack on an adenine of the upstream intron. The attack takes place by a hydroxyl group of an end of the downstream intron. And then a circular intron is formed.(Figure 2)<br> |
<img src="https://static.igem.org/mediawiki/2014/2/2b/SS2.png" width="600px"></img><br> | <img src="https://static.igem.org/mediawiki/2014/2/2b/SS2.png" width="600px"></img><br> | ||
<b>Fig.3 Self-splicing in T4 phage: the third step (Blue: intron, Orange: exon)</b><br><br> | <b>Fig.3 Self-splicing in T4 phage: the third step (Blue: intron, Orange: exon)</b><br><br> | ||
</p> | </p> | ||
<h2>The permuted intron-exon method: PIE method</h2> | <h2>The permuted intron-exon method: PIE method</h2> | ||
− | <p>Two exons are connected with each other in the circularization system; furthermore an exon can theoretically be circularized by the system. (Fig.4)<br> | + | <p> Two exons are connected with each other in the circularization system; furthermore an exon can theoretically be circularized by the system. (Fig.4)<br> |
<img src="https://static.igem.org/mediawiki/2014/a/a1/PIEGIFU3.png" width="650px"></img><br><br> | <img src="https://static.igem.org/mediawiki/2014/a/a1/PIEGIFU3.png" width="650px"></img><br><br> | ||
<img src="https://static.igem.org/mediawiki/2014/d/d2/PIEGIFU2.png" width="600px"></img><br> | <img src="https://static.igem.org/mediawiki/2014/d/d2/PIEGIFU2.png" width="600px"></img><br> |
Revision as of 02:20, 2 September 2015