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Team:Tsinghua/Experiment2
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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. | 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. | ||
| - | [[Image:J61031.jpg|800px|J61031|center]] | + | [[Image:J61031.jpg|800px|J61031|center|thumb|The cosmid map]] |
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[[Image:cosmidcostruct.jpg|800px|cosmidcostruct|center]] | [[Image:cosmidcostruct.jpg|800px|cosmidcostruct|center]] | ||
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=Characterization of Therapeutic DNA= | =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. | 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. | ||
Revision as of 16:44, 21 October 2009
| Home | Background | Brainstorming | Design | Experiment | Results | Conclusion | Protocol |
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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.
