Difference between revisions of "Part:BBa K1477030"

 
(→‎Characterization and Applications of BBa_K1477030)
 
(4 intermediate revisions by 2 users not shown)
Line 1: Line 1:
−
 
 
__NOTOC__
 
__NOTOC__
 
{|width='80%'
 
{|width='80%'
−
|-
+
 
−
|width=35% valign='top'|
+
−
[[Image:Terminator.png]]
+
−
<partinfo>BBa_K1477030 parameters</partinfo>
+
−
| width=50% valign='top' style='border: 1px solid black'|
+
 
<partinfo>BBa_K1477030 short</partinfo>
 
<partinfo>BBa_K1477030 short</partinfo>
−
* T7 inducable promoter and an Endolysin terminater in-vitro used to recognize specific virus's, when the virus is present the cells lyse.  
+
* T7 strong promoter coupled with an endolysin generator designed to short circuit the replication cycle of T7 bacteriophage.
−
 
+
<partinfo>BBa_K1477030 SequenceAndFeatures</partinfo>  
−
<hr>
+
−
<partinfo>BBa_K1477030 SpecifiedComponents</partinfo>
+
−
|}
+
−
 
+
−
'''Secondary Structure'''
+
−
 
+
−
[[Image:Mfold-K1477030-1.png]]
+
  
−
<hr>
+
===Characterization and Applications of BBa_K1477030===
−
'''Measurement'''
+
<p>
−
* [http://openwetware.org/wiki/Cconboy:Terminator_Characterization/Results How these parts were measured]
+
[[File:Electrophoresis_K1477030.png|200px|thumb|left|alt text]]
 +
</p>
 +
<p>
 +
In correspondence with a 500 basepair DNA ladder, our part measures at ~2500 base pairs which corresponds with the sum of the BioBricks and the pSB1C3 backbone. Plasmid DNA isolated from two subclones
 +
</p>
 +
<p>
 +
[[File:Coliphage_lysis_of_BBa_K1477030.png|400px|thumb|left|alt text]]
 +
</p>
 +
<p>
 +
TEST FOR IMMEDIATE LYSIS OF TOP10 CELLS TRANSFORMED WITH BBa_K1477030.
 +
This immediate-lysis device was designed to be an immediate lysis cassette for T7 infection of transformed Top10 cells. The expected result was that a culture of transformed cells would be resistant to infection by T7 phage but not by an unrelated phage.  This experiment represents transformed cells infected at a ratio of 100 cells to 1 pfu T7 phage.  If functioning, the immediate lysis cassette should have stopped the propagation of T7 infection.  What may be lacking is the presence of holin that should increase the effectiveness of the endolysin.
 +
</p>

Latest revision as of 01:27, 3 November 2014

T7 strong promoter + endolysin cassette
  • T7 strong promoter coupled with an endolysin generator designed to short circuit the replication cycle of T7 bacteriophage.

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 345
    Illegal AgeI site found at 415
  • 1000
    COMPATIBLE WITH RFC[1000]

Characterization and Applications of BBa_K1477030

alt text

In correspondence with a 500 basepair DNA ladder, our part measures at ~2500 base pairs which corresponds with the sum of the BioBricks and the pSB1C3 backbone. Plasmid DNA isolated from two subclones

alt text

TEST FOR IMMEDIATE LYSIS OF TOP10 CELLS TRANSFORMED WITH BBa_K1477030. This immediate-lysis device was designed to be an immediate lysis cassette for T7 infection of transformed Top10 cells. The expected result was that a culture of transformed cells would be resistant to infection by T7 phage but not by an unrelated phage. This experiment represents transformed cells infected at a ratio of 100 cells to 1 pfu T7 phage. If functioning, the immediate lysis cassette should have stopped the propagation of T7 infection. What may be lacking is the presence of holin that should increase the effectiveness of the endolysin.