Team:Tsinghua/Experiment2

=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.