Difference between revisions of "Part:BBa K1943020"
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<partinfo>BBa_K1943020 short</partinfo> | <partinfo>BBa_K1943020 short</partinfo> | ||
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+ | GECO family is a series of fluorescent proteins that artificially engineered to response to changes in the concentration of calcium ion Ca2<sup>+</sup>. They can help to visualize intracellular signaling activity by the intensity. According to Zhao’s article, there is optimally 110-folds of emission change. | ||
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+ | By recombining the GCaMPs and GFP, firstly they got the G-GECO1(green). The indicator we used is R-GECO1 derived from mApple and G-GECO1.1. The results of direct evolution also included GEM-GECO and Blue GECO. (see the genealogy of GECO series ) | ||
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+ | [[File:T--SUSTech Shenzhen--geco.jpg]] | ||
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+ | These various indicators is helpful in live cell image, and improve Ca2+ imagining in neurons and muscles as well. In our experiment, R-GECO1 was used to detect cell response to mechanical vibration. | ||
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+ | There have been an iGEM team submitted its sequence in 2012 (<partinfo>BBa_K881000</partinfo>). But, unfortunately, it did not meet the RFC10 standard because of the restriction sites in its original sequence. For the convenience, we mutated these regions to remove the repeated cloning sites. | ||
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+ | [[File:T--SUSTech Shenzhen--geco_1.jpg]] | ||
Mutated R-GECO CDS without 2 PstI in original sequence. | Mutated R-GECO CDS without 2 PstI in original sequence. |
Revision as of 18:07, 21 October 2016
R-GECO
GECO family is a series of fluorescent proteins that artificially engineered to response to changes in the concentration of calcium ion Ca2+. They can help to visualize intracellular signaling activity by the intensity. According to Zhao’s article, there is optimally 110-folds of emission change.
By recombining the GCaMPs and GFP, firstly they got the G-GECO1(green). The indicator we used is R-GECO1 derived from mApple and G-GECO1.1. The results of direct evolution also included GEM-GECO and Blue GECO. (see the genealogy of GECO series )
These various indicators is helpful in live cell image, and improve Ca2+ imagining in neurons and muscles as well. In our experiment, R-GECO1 was used to detect cell response to mechanical vibration.
There have been an iGEM team submitted its sequence in 2012 (BBa_K881000). But, unfortunately, it did not meet the RFC10 standard because of the restriction sites in its original sequence. For the convenience, we mutated these regions to remove the repeated cloning sites.
Mutated R-GECO CDS without 2 PstI in original sequence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 914
Illegal SapI.rc site found at 111
Illegal SapI.rc site found at 820