Difference between revisions of "Part:BBa K1362000"
Line 2: Line 2:
 
<partinfo>BBa_K1362000 short</partinfo>
 
<partinfo>BBa_K1362000 short</partinfo>
  
−
???
 
  
 +
[[file:Heidelberg2014_BBa_K1362000.png|600px|'''Figure 1''': NpuDnaE intein RFC [i] circularization construct]]
  
−
===Usage and Biology===
+
===Circular Proteins===
  
−
https://static.igem.org/mediawiki/2014/9/9b/SampleData_Circularization.png
+
[[File:Heidelberg2014_circ_lamlys.png|400px|thumb|right|'''Figure 2''': The 3D structure of linear lambda lysozyme (purple) is compared with a model predicted by modeller (ref?) of circular lambda lysozyme (yellow) with a rigid linker (green)]]
 +
 
 +
===Usage===
 +
 
 +
[[File:Heidelberg2014_SampleData_Circularization.png|400px|thumb|left|'''Figure 3''': In a single step the protein of interest is cloned into the split intein circularization construct (1.). The resulting plasmid (2.) can be used to express a fusion protein (3.). In an autocatalytic in vivo reaction, the circular protein (4.) is formed.]]
 +
 
 +
===Results===
 +
 
 +
[[File:Heidelberg2014_WB_36.jpg|400px|thumb|right|'''Figure 4''': Western Blot to prove circularization.
 +
In order to test whether lambda lysozyme is circular, we did a Western Blot with Penta·His Antibody. There is a clear shift between linear and circular samples.]]
 +
 
 +
[[placeholder|400px|thumb|right|'''Figure 5''': xyla-wb]]
  
 
<!-- -->
 
<!-- -->

Revision as of 15:13, 17 October 2014

NpuDnaE intein RFC[105] circularization construct


Figure 1: NpuDnaE intein RFC [i] circularization construct

Circular Proteins

Figure 2: The 3D structure of linear lambda lysozyme (purple) is compared with a model predicted by modeller (ref?) of circular lambda lysozyme (yellow) with a rigid linker (green)

Usage

Figure 3: In a single step the protein of interest is cloned into the split intein circularization construct (1.). The resulting plasmid (2.) can be used to express a fusion protein (3.). In an autocatalytic in vivo reaction, the circular protein (4.) is formed.

Results

Figure 4: Western Blot to prove circularization. In order to test whether lambda lysozyme is circular, we did a Western Blot with Penta·His Antibody. There is a clear shift between linear and circular samples.

400px|thumb|right|Figure 5: xyla-wb

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 931
    Illegal AgeI site found at 1043
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1220
    Illegal BsaI.rc site found at 145