Difference between revisions of "Part:BBa K1062004:Experience"

Revision as of 13:20, 6 September 2018

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Applications of BBa_K1062004

User Reviews

UNIQb5e1df35e3c89939-partinfo-00000000-QINU

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USTC_China iGEM13

In 2018 OUC-China's project, we used csy4 as a main role in our toolkit. We design a toolkit focused on translational regulation, which is composed of a RNA endoribonuclease (Csy4) and a RNA module (hairpin). Csy4 (Csy6f), a member of CRISPR family, recognizes a specific 16-nt nucleotide repetitive sequence(hairpin). The RNA module was constructed by inserting the 16nt Csy4 recognition site between a RBS and cis-repressive RNA element, which can be specifically cleaved upon Csy4 expression, so that the RBS is masked. Cleaved at the specific recognition site, it can release the masked RBS, thus endowing the programming of gene expression in the translation level with higher feasibility.

We found that csy4 is a perfect part for our toolkit. Because once the RNA/Csy4 complex formed, the structure become really stable and hard to separate which make our toolkit stable when it function.

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 <I>USTC_China iGEM13</I>
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-In USTC_China iGEM13's project,we bulit the N-terminal TD1 modified GFP to make it is capable of transporting through skin barrier, then induced expression in BL21 and Bacillus Subtilis WB800N with IPTG and tested positive by SDS-PAGE and ultraviolet ultraviolet spectrometry. Students in Wen lab validated the efficient transdermal function of N-terminal TD1 modified GFP(not BBa_E0040 ).
+In 2018 OUC-China's project, we used csy4 as a main role in our toolkit. We design a toolkit focused on translational regulation, which is composed of a RNA endoribonuclease (Csy4) and a RNA module (hairpin). Csy4 (Csy6f), a member of CRISPR family, recognizes a specific 16-nt nucleotide repetitive sequence(hairpin). The RNA module was constructed by inserting the 16nt Csy4 recognition site between a RBS and cis-repressive RNA element, which can be specifically cleaved upon Csy4 expression, so that the RBS is masked. Cleaved at the specific recognition site, it can release the masked RBS, thus endowing the programming of gene expression in the translation level with higher feasibility.
-[[Image:USTC_China_iGEM13_BBa_K1074006_1.png|thumb|left|800px|figure 1 SDS-PAGE of N-terminal TD1 modified GFP]]
-[[Image:USTC_China_iGEM13_BBa_K1074006_2.JPG|thumb|left|600px|figure 2 Fluorescence detection of N-terminal TD1 modified GFP expression with and without IPTG induction]]
+We found that csy4 is a perfect part for our toolkit. Because once the RNA/Csy4 complex formed, the structure become really stable and hard to separate which make our toolkit stable when it function.
-[[Image:USTC_China_iGEM13_BBa_K1074000_4.jpg|thumb|left|600px|figure 3 Our Transdermal test device]]
-[[Image:USTC_China_iGEM13_BBa_K1074006_3.JPG|thumb|left|600px|figure 4 Results of transdermal test of N-terminal TD1 modified GFP and  unmodified GFP]]