Team:TU-Munich/Results/KillSwitch

From 2013.igem.org

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[[File:TUM13_PartsRegistryKillSwitch.png|thumb|center|750px| '''Figure 1''': [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159118 Registry entry] of the PhyB/PIF6 version of the Kill Switch, BBa_K1159118]]
[[File:TUM13_PartsRegistryKillSwitch.png|thumb|center|750px| '''Figure 1''': [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159118 Registry entry] of the PhyB/PIF6 version of the Kill Switch, BBa_K1159118]]
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==Transformation of the large DNA constructs==
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moss -> no homologuos regions
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bacteria -> codon usage
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==Level of promoter activity==
[[File:PBI_376_f1.gif|thumb|right|500px| '''Figure 2''': Comparative expression performance of different constitutive mammalian and plant promoters in Physcomitrella patens, [[http://www.ncbi.nlm.nih.gov/pubmed/19021876 Gitzinger et al., 2009]]]]
[[File:PBI_376_f1.gif|thumb|right|500px| '''Figure 2''': Comparative expression performance of different constitutive mammalian and plant promoters in Physcomitrella patens, [[http://www.ncbi.nlm.nih.gov/pubmed/19021876 Gitzinger et al., 2009]]]]
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==Photosensitivity==
[[File:TUM13_Foto_Labpics2.jpg|thumb|right|500px| '''Figure 3''': Working with the photosensitive protoplasts]]
[[File:TUM13_Foto_Labpics2.jpg|thumb|right|500px| '''Figure 3''': Working with the photosensitive protoplasts]]
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* in bakkis nicht exprimiert [codon usage]
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* promotor zu stark [bild vgl prom stärken]
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* konstrukt zu groß für trafo [homologe regionen??]
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* filter nicht optimal [bild filter, ingmar @ blaulicht bench?]
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==References==
==References==

Revision as of 22:06, 28 October 2013


Kill Switch

The Kill Switch mechanism is the most complex and ambitious aspect of our project on the protein level. The DNA constructs are very large and even exceed the Part Registry´s frame (Fig. 1). For details on its design and function see here. We were very excited to start experimenting with the killswitch moss. However, when we opened the redlight filter foil, we found none of the allegedly transgenic moss lines had survived. There is a variety of possible reasons for this outcome that need to be discussed in order to improve our approach.

Figure 1: [http://parts.igem.org/wiki/index.php?title=Part:BBa_K1159118 Registry entry] of the PhyB/PIF6 version of the Kill Switch, BBa_K1159118

Transformation of the large DNA constructs

moss -> no homologuos regions bacteria -> codon usage

Level of promoter activity

File:PBI 376 f1.gif
Figure 2: Comparative expression performance of different constitutive mammalian and plant promoters in Physcomitrella patens, http://www.ncbi.nlm.nih.gov/pubmed/19021876 Gitzinger et al., 2009

Photosensitivity

Figure 3: Working with the photosensitive protoplasts

[http://www.plant-biotech.net/paper/CurrGenet_2003_hohe.pdf Hohe et al., 2003] "Targeted gene-knockout in Physco" paper

Figure 3: Working with the photosensitive protoplasts
Figure 3: Working with the photosensitive protoplasts
Figure 3: Working with the photosensitive protoplasts




References

Retrieved from "http://2013.igem.org/Team:TU-Munich/Results/KillSwitch"