Difference between revisions of "Team:Alverno CA/Design"
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<h1><center>Design</center></h1> | <h1><center>Design</center></h1> | ||
| − | <p> | + | <p>The center of of our project is to counteract a problem in synthetic biology known as supercoiling. Supercoiling is a possible negative side effect that can occur in the construction of a multi-genetic circuit. When the two genes transcribe, the double helix is opened in order to allow for the expression for the genes, however, the dna in between the two genes receive extraordinary amounts of tension when both genes try to transcribe at the same time, which leads to the over transcription of one gene, and the under expression of the other gene . This can greatly affect the predictability of gene expression. </p> |
| + | <p> In order to solve this problem, we have designed a plasmid where two genes—RFP and GFP—are constructed onto the same plasmid. We then tested three different methods in order to eliminate the appearance of supercoiling. The first method we tried was adding random base pair sequence in between the two genes, with lengths ranging in between fifty and one thousand and five hundred base pairs. We created this in the hope that the space in between the two genes would alleviate the tension that causes the supercoils. Another of our methods was to use a Dcas9 clamp on the space between the two base pairs to hopefully lessen the effect of supercoiling. We utilized Golden Gate Assembly in order easily build several variations of these plasmids by cloning with different parts. <p> | ||
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Revision as of 21:13, 15 October 2016
Design
The center of of our project is to counteract a problem in synthetic biology known as supercoiling. Supercoiling is a possible negative side effect that can occur in the construction of a multi-genetic circuit. When the two genes transcribe, the double helix is opened in order to allow for the expression for the genes, however, the dna in between the two genes receive extraordinary amounts of tension when both genes try to transcribe at the same time, which leads to the over transcription of one gene, and the under expression of the other gene . This can greatly affect the predictability of gene expression.
In order to solve this problem, we have designed a plasmid where two genes—RFP and GFP—are constructed onto the same plasmid. We then tested three different methods in order to eliminate the appearance of supercoiling. The first method we tried was adding random base pair sequence in between the two genes, with lengths ranging in between fifty and one thousand and five hundred base pairs. We created this in the hope that the space in between the two genes would alleviate the tension that causes the supercoils. Another of our methods was to use a Dcas9 clamp on the space between the two base pairs to hopefully lessen the effect of supercoiling. We utilized Golden Gate Assembly in order easily build several variations of these plasmids by cloning with different parts.
