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Team:Tsinghua/Experiment2
From 2009.igem.org
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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.
Characterization of Therapeutic DNA
The characterization of Therapeutic DNA is mainly via fluorescent observation and co-functioning testing together with the synthesized GenSniper genome. If the experiments proceed as expected, we will also identify whether Therapeutic DNA has been packaged into the GenSniper genome via PCR.
