Difference between revisions of "Part:BBa K1033932:Experience"
(→‎Characterization)
 
(6 intermediate revisions by 4 users not shown)
Line 5: Line 5:
  
 
===Applications of BBa_K1033932===
 
===Applications of BBa_K1033932===
 +
===User review of SHSBNU_China===
 +
[[File: SHSBNU 17 40aL1.jpg|600px|thumb|center|Figure 1]]
 +
 +
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).
 +
 +
[[File: SHSBNU 17 40aL2.jpg|600px|thumb|center|Figure 2]]
 +
 +
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.
 +
 +
[[File: SHSBNU 17 40aL3.jpg|600px|thumb|center|Figure 3]]
 +
 +
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===
 
===User Reviews===
 
<!-- DON'T DELETE --><partinfo>BBa_K1033932 StartReviews</partinfo>
 
<!-- DON'T DELETE --><partinfo>BBa_K1033932 StartReviews</partinfo>
−
<!-- Template for a user review
+
 
 
{|width='80%' style='border:1px solid gray'
 
{|width='80%' style='border:1px solid gray'
 
|-
 
|-
 
|width='10%'|
 
|width='10%'|
−
<partinfo>BBa_K1033932 AddReview number</partinfo>
+
<partinfo>BBa_K1033932 AddReview 5</partinfo>
 
<I>SUSTech_Shenzhen 2016</I>
 
<I>SUSTech_Shenzhen 2016</I>
 
|width='60%' valign='top'|
 
|width='60%' valign='top'|
−
SUSTech_Shenzhen 2016 use this part to construct a new composite part, which could work as a reporter. When cultured in LB, the expression of color seemed not very obvious.
+
SUSTech_Shenzhen 2016 use this part to construct two new composite parts, <partinfo>BBa_K1943006</partinfo> and <partinfo>BBa_K1943007</partinfo>  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.
 +
https://static.igem.org/mediawiki/parts/e/e1/SUSTech_Shenzhen-SUSTecho.png
 
|};
 
|};
−
 
−
<!-- DON'T DELETE --><partinfo>BBa_K1033932 EndReviews</partinfo>
 

Latest revision as of 13:33, 12 October 2019


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1033932

User review of SHSBNU_China

Figure 1

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

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

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

UNIQa2ea93ca3844b857-partinfo-00000000-QINU

•••••

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. SUSTech_Shenzhen-SUSTecho.png

;