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Team:Tsinghua/Experiment
From 2009.igem.org
| Background | Brainstorming | Design | Experiment | Results | Notebook |
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Contents |
Synthesis of the Therapeutic DNA
In this part, we aims at constructing a molecular cloning vector with a cos site which enables it to be packaged into the gene therapy vector- in other words, to construct a cosmid. In order to detect the expression of the therapeutic DNA, we construct this cosmid based on the scanfold of Parts-J61031 encoding a RFP-expressing segment.
This cosmid mainly consists of the following segments:
1) origin of replication: we insert O gene and P gene from bacteriophage lambda into J61031, which are resoonsible for the late phase replication and package of the wild type circular bacteriophage lambda genome
2) cos site: necessary for the specific package of the Therapeutic DNA into the gene therapy vector.
3) RFP-expressing segment: originally integrated into the plamid of J61031.
Synthesis of the Gene Therapy Production System
Bottom-Up Approach
In the bottom-up approach, we amplify the biobricks from both the bacteriophage lambda and the adenovirus genome and incorporate them with a given order into molecular cloning vector(s).
Specifically, we choose two molecular cloning vectors to encode two sections of the gene therapy vector genome, pET28a and pACYCDuet1.
