Difference between revisions of "Part:BBa K2384013"
 
(2 intermediate revisions by one other user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K2384013 short</partinfo>
 
<partinfo>BBa_K2384013 short</partinfo>
  
−
<p>Xylose-inducible expression system: strong<b> P<i>xylA</i></b> promoter and repressor gene <b><i>xylR</i></b>; after xylose addition, the repressor is released from the P<i>xylA</i> that activates transcription initiation.([[Part:BBa_K733002|BBa_K733002]]) They have been designed for high expression in <i>B. megaterium</i> This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell([[Part:BBa_K1460002|BBa_K1460002]]). This year our parts were doing codon optimisation but without terminator codon.</p>
+
<p>Xylose-inducible expression system: strong<b> P<i>xylA</i></b> promoter and repressor gene <b><i>xylR</i></b>; after xylose addition, the repressor is released from the P<i>xylA</i> that activates transcription initiation.([[Part:BBa_K733002|BBa_K733002]]) They have been designed for high expression in <i>B. megaterium</i>. This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell([[Part:BBa_K1460002|BBa_K1460002]]). This year our parts were doing codon optimisation but without terminator codon.</p>
  
  
Line 14: Line 14:
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--GST-CRS51.png|thumb|400px|'''After Codon Adjustment''']]</th>
+
<th>[[Image:T--FAFU-CHINA--GST-CRS51.png|thumb|400px|'''Figure 1:After Codon Adjustment''']]</th>
 
</tr></table>
 
</tr></table>
  
  
−
3.GC Content:
+
2.GC Content:
 
The comparison of  GC content between original sequence and optimized sequence
 
The comparison of  GC content between original sequence and optimized sequence
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--GST-CRS52.png|thumb|400px|'''After Optimization''']]</th>
+
<th>[[Image:T--FAFU-CHINA--GST-CRS52.png|thumb|400px|'''Figure 2:After Optimization''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--GST-CRS5 SDS PAGE.jpg|thumb|400px|'''The expression of GST-CRS5 detected using SDS-PAGE ''']]</th>
+
<th>[[Image:T--FAFU-CHINA--GST-CRS5 SDS PAGE.jpg|thumb|400px|'''Figure 3:The expression of GST-CRS5 detected using SDS-PAGE ''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--M-GST-CRS5.jpg|thumb|400px|'''Plasmid digested by EcoRI and HindIII ''']]</th>
+
<th>[[Image:T--FAFU-CHINA--M-GST-CRS5.jpg|thumb|400px|'''Figure 4:Plasmid digested by EcoRI and HindIII ''']]</th>
 
</tr></table>
 
</tr></table>
  
 
<table><tr>
 
<table><tr>
−
<th>[[Image:T--FAFU-CHINA--GST-CRS5-Toggle-switch.png|thumb|400px|'''Toggle switch ''']]</th>
+
<th>[[Image:T--FAFU-CHINA--GST-CRS5-Toggle-switch.png|thumb|400px|'''Figure 5:Our genetic circuit ''']]</th>
 
</tr></table>
 
</tr></table>
  

Latest revision as of 02:01, 2 November 2017


Repressor gene xylR+Strong PxylA promoter+RBS+GST-CRS5+His-tag

Xylose-inducible expression system: strong PxylA promoter and repressor gene xylR; after xylose addition, the repressor is released from the PxylA that activates transcription initiation.(BBa_K733002) They have been designed for high expression in B. megaterium. This part serves as a functional MT for heavy metal binding and conferred resistance toxic metal cadmium. GST as a fusion partenr is to enhance stability of expression.The original part was from Cornell(BBa_K1460002). This year our parts were doing codon optimisation but without terminator codon.


Usage and Biology

Optimization Report

1.Codon Used Adjustment The best value is 1 for sequence optimization.

Figure 1:After Codon Adjustment


2.GC Content: The comparison of GC content between original sequence and optimized sequence

Figure 2:After Optimization
Figure 3:The expression of GST-CRS5 detected using SDS-PAGE
Figure 4:Plasmid digested by EcoRI and HindIII
Figure 5:Our genetic circuit

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 2159
    Illegal XhoI site found at 847
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 1571