Difference between revisions of "Part:BBa K1033932:Experience"
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| − | <!-- DON'T DELETE --><partinfo> | + | ===Characterization=== |
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| + | <partinfo>BBa_K1033910 AddReview 3</partinfo> | ||
| + | <I>NWU-China 2019</I> | ||
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| + | The NWU-China team overexpressed spisPink protein in E. coli and extracted and purified. The maximum absorbance was measured in a 96-well plate using a microplate reader (Synergy 2). | ||
| + | https://2019.igem.org/wiki/images/7/78/T--NWU-China--characterization-BBa_K1033932%28spisPink%29.png | ||
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Revision as of 15:51, 9 October 2019
This experience page is provided so that any user may enter their experience using this part.
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how you used this part and how it worked out.
Applications of BBa_K1033932
User review of SHSBNU_China
Figure 1 TtrS/R system and ThsS/R system ThsS (BBa_K2507000) and ThsR (BBa_K2507001), both codon-optimized for E. coli, are two basic parts which belong to the two-component system from the marine bacterium Shewanella halifaxensis. By linking thsR with chromoprotein genes (BBa_K2507009, BBa_K2507010, BBa_K2507011) or the violacein producing operon vioABDE (BBa_K2507012), this system can respond to thiosulfate by producing a signal visible to the naked eye, such as chromoproteins (spisPink-pink chromoprotein, gfasPurple-purple chromoprotein, amilCP-blue chromoprotein) or a dark-green small-molecule pigment (protoviolaceinic acid). E. coli-codon-optimized TtrS(BBa_K2507002) and TtrR (BBa_K2507003) are two basic parts which are derived from the two-component system of the marine bacterium Shewanella baltica.
By linking ttrR with chromoprotein genes (BBa_K2507009, BBa_K2507010, BBa_K2507011) or the violacein producing operon vioABDE (BBa_K2507012), this system can respond to thiosulfate by producing a signal visible to the naked eye such as chromoproteins (spisPink-pink chromoprotein, gfasPurple-purple chromoprotein, amilCP-blue chromoprotein) or a dark-green small-molecule pigment (protoviolaceinic acid).
Figure 2. Characterization of the ThsS/R and TtrS/R system by observing the chromprotein expression levels. We added 1mM, 0.1mM, 0.01mM and 0 Na2S2O3 to ThsS/R system and added 2.5mM, 1mM, 0.1mM and 0 Na2S4O6·2H2O to ThsS/R system. The results demonstrate there is an obvious response in the ThsS/R system. In a. gfasPurple system, the response curve is obvious, and b. spisPink & c. amilCP with rather heavy leaky expression without inducer. d.e.f. However, in TtrS/R system there is no clear result.
Figure 3. Characterization of the ThsS/R and TtrS/R system by observing the protoviocaceinic acid( dark-green pigment). We added 1mM, 0.1mM, 0.01mM and 0 Na2S2O3 to ThsS/R system and added 2.5mM, 1mM, 0.1mM and 0 Na2S4O6·2H2O to ThsS/R system. The results demonstrate there is an obvious response in the ThsS/R system with rather heavy leaky expression without inducer. And the Ths/R+protoviolaceinic acid works rather well!
User Reviews
UNIQ67aebfe582b4aafe-partinfo-00000000-QINU
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SUSTech_Shenzhen 2016 |
SUSTech_Shenzhen 2016 use this part to construct two new composite parts, BBa_K1943006 and BBa_K1943007 which could work as a reporter. When cultured in LB, the expression of color with the help of strong promoter-RBS combination gave excellent effect. While the one with weaker promoter-RBS combination did not work very well.
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Characterization
UNIQ67aebfe582b4aafe-partinfo-00000004-QINU
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NWU-China 2019 |
The NWU-China team overexpressed spisPink protein in E. coli and extracted and purified. The maximum absorbance was measured in a 96-well plate using a microplate reader (Synergy 2).
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