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Team:Tsinghua/Result2
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GuoQiangChen (Talk | contribs) (→Co-Functioning of Therapeutic DNA with GenSniper Genome) |
GuoQiangChen (Talk | contribs) (→Co-Functioning of Therapeutic DNA with GenSniper Genome) |
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[[Image:lambda+cos.png|600px|center|thumb|lambda+cos,A1/3K1/3C]] | [[Image:lambda+cos.png|600px|center|thumb|lambda+cos,A1/3K1/3C]] | ||
| - | [[Image:electrocos.png|600px|center|electrocos]] | + | The next crucial step is that Theraputic DNA can be packaged into the GenSniper genome. As showed in the results of Module I, we have gained the desired the GenSniper viroin without Therapeutic DNA packaging. When we further co-tranformed the GenSniper genome and Therapeutic DNA for GenSniper viroin production with the packaged DNA, the purified solution also contains the desired protein complex. |
| - | [[Image:electro0.png|600px|center|electro0]] | + | |
| + | In addition, PCR identification of cos site indicates positive result. | ||
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| + | [[Image:electrocos.png|600px|center|electrocos|thumb|Electronic spectroscopy picture of the GenSniper viroin with Therapeutic DNA package and without Targeted Biobrick modification]] | ||
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| + | [[Image:electro0.png|600px|center|electro0|thumb|Electronic spectroscopy picture of the control group]] | ||
[[Image:cosPCR.png|600px|center|cosPCR]] | [[Image:cosPCR.png|600px|center|cosPCR]] | ||
Revision as of 18:16, 18 October 2009
Construction and Characterization of Therapeutic DNA
The map of the Therapeutic DNA is listed above. It contains a incorporated cos site, O and P genes for replication and RFP expression segment. We should note that the RFP expression module in this Therapeutic DNA is downstream a prokaryotic promoter, which enable us to trace the function of the Therapeutic DNA according to red fluorescence. However, when applied to clinical use, the RFP expression segment can be replaced by the real therapeutic genes.
We have completed the construction of Therapeutic DNA and proved that it contains the cos site for the package into the GenSniper viroin through PCR.
In theory, the therapeutic DNA should enable the bacteria containing it to express RFP. Fortunately, the red fluorescent was so strong to be observed via naked eyes. Fluorescence microscopy also confirmed our observation.
Since the original Part_J61031 contains both Kan and Amp resistance, which will render the incompatibility for Therapeutic DNA and pET-28a-based constructs, we moecularly cutted the region of Kan-Amp resistance and reconstructed a cosmid with only Amp resistance gene. B shows the plate with Amp and Therapeutic DNA transformed E.coli. C shows the plate with Kan and Therapeutic DNA transformed E.coli. These results suggests that the single resistance "cosmid" has been successfully constructed.
Co-Functioning of Therapeutic DNA with GenSniper Genome
The next crucial step is that Theraputic DNA can be packaged into the GenSniper genome. As showed in the results of Module I, we have gained the desired the GenSniper viroin without Therapeutic DNA packaging. When we further co-tranformed the GenSniper genome and Therapeutic DNA for GenSniper viroin production with the packaged DNA, the purified solution also contains the desired protein complex.
In addition, PCR identification of cos site indicates positive result.
