Difference between revisions of "Part:BBa S04615"
Line 19: Line 19:
  
 
<div style="color:black;font-size:25px">Improvement by SDU_CHINA 2019</div>
 
<div style="color:black;font-size:25px">Improvement by SDU_CHINA 2019</div>
−
In 2018, the SDU-CHINA team optimized the BBa_K3127995 part to reduce its leakage under the red light. We have improved its promoters this year, which has further bettered the performance of this component.
+
 
 +
In 2018, the SDU-CHINA team optimized the K592001 part to reduce its leakage under the red light. We have improved its promoters this year, which has further bettered the performance of this component.
 +
 
 +
The component BBa_K2627000 proved the progress of the optimized CcaS by sufficient characterization, so we directly replaced K592001 in BBa_S04615 with the optimized K2627000 when characterizing this component. In 2014, Schmidl S R , Sheth R U, Wu A, et al. optimized CcaS by constructing an RBS library. This document has fully proven that the performance of components with optimized RBS is better than S04615. In 2019, our team SDU-CHINA has further improved the components based on them. We have introduced site-directed mutagenesis to the improved promoter, further bettering the performance based on all previous studies.
 +
==Results==
 +
===Characaterization of BBa_K2627000===
 +
<p>The plasmid carrying our RBS is called 5-10#10.  There is less fluorescence intensity than #10 under light of 640-650 nm wavelength.<br>
 +
The following figure shows the relative fluorescence intensity induced by the light of 640-650 nm wavelength(green light) and the light of 530-535 nm wavelength(red light).</p>
 +
 
 +
<div style="display:flex;justify-content:space-around">[[File:T-SDU CHINA-BW25113--10.png|300px|thumb|center|<b>Figure 1:</b> Characterization of this old part in E.coli BW25113 in M9 medium whose carbon source is glucose.]][[File:T-SDU CHINA-BW25113-5-10-10.png|300px|thumb|center|<b>Figure 2:</b> Characterization of this improved part in E.coli BW25113 in M9 medium whose carbon source is glucose.]]</div>
 +
<div style="display:flex;justify-content:space-around">[[File:T-SDU CHINA-TOP10--10.png|300px|thumb|center|<b>Figure 3:</b> Characterization of this old part in E.coli TOP10 in M9 medium whose carbon source is glucose.]][[File:T-SDU CHINA-TOP10-5-10-10.png|300px|thumb|center|<b>Figure 7:</b> Characterization of this improved part in E.coli TOP10 in M9 medium whose carbon source is glucose.]]</div>

Revision as of 17:38, 20 October 2019


B0034:K592001

Construction intermediate

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 624
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2270
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1288
  • 1000
    COMPATIBLE WITH RFC[1000]


Improvement by SDU_CHINA 2019

In 2018, the SDU-CHINA team optimized the K592001 part to reduce its leakage under the red light. We have improved its promoters this year, which has further bettered the performance of this component.

The component BBa_K2627000 proved the progress of the optimized CcaS by sufficient characterization, so we directly replaced K592001 in BBa_S04615 with the optimized K2627000 when characterizing this component. In 2014, Schmidl S R , Sheth R U, Wu A, et al. optimized CcaS by constructing an RBS library. This document has fully proven that the performance of components with optimized RBS is better than S04615. In 2019, our team SDU-CHINA has further improved the components based on them. We have introduced site-directed mutagenesis to the improved promoter, further bettering the performance based on all previous studies.

Results

Characaterization of BBa_K2627000

The plasmid carrying our RBS is called 5-10#10. There is less fluorescence intensity than #10 under light of 640-650 nm wavelength.
The following figure shows the relative fluorescence intensity induced by the light of 640-650 nm wavelength(green light) and the light of 530-535 nm wavelength(red light).

Figure 1: Characterization of this old part in E.coli BW25113 in M9 medium whose carbon source is glucose.
Figure 2: Characterization of this improved part in E.coli BW25113 in M9 medium whose carbon source is glucose.
Figure 3: Characterization of this old part in E.coli TOP10 in M9 medium whose carbon source is glucose.
Figure 7: Characterization of this improved part in E.coli TOP10 in M9 medium whose carbon source is glucose.