Difference between revisions of "Part:BBa K2350021"
(→‎Functional Test)
 
(26 intermediate revisions by one other user not shown)
Line 3: Line 3:
 
<partinfo>BBa_K2350021 short</partinfo>
 
<partinfo>BBa_K2350021 short</partinfo>
  
−
===Endolysin-Holin-NrtA===
+
===Holin-Endolysin-NrtA===
−
 
+
　　In this part, we combine holin (BBa_K2350019), endolysin (BBa_K2350020), and NrtA (BBa_K2350004). NrtA protein can stick to periplasmic membrane through a flexible linker to capture nitrite or nitrate in the periplasm. Therefore, we make use of NrtA in nitrogen starvation in our 2017 NYMU project. After we reach our goal, we need to kill E.coli to prevent contamination. When the lactose is added into the environment, the suicide mechanism, holin and endolyin, is induced. For more detail information, please see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, BBa_K2350019 for holin part, and BBa_K2350020 for endolysin-holin part.
  
  
 
===Promoter, RBS, and Terminator===
 
===Promoter, RBS, and Terminator===
 +
　　For more details of the connected promoter, RBS, and terminator, you can see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, and BBa_K2350019 for holin part.
  
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
  
−
<!-- -->
+
===Result===
−
<span class='h3bb'>Sequence and Features</span>
+
　　Figure 1 is restriction enzyme check electrophoresis result of Holin-Endolysin-NrtA construct. We use XmaI to digest sample 1~3. The result shows that 2 and 3 are right. M represents 1 kb marker.
−
<partinfo>BBa_K2350021 SequenceAndFeatures</partinfo>
+
  
  
−
<!-- Uncomment this to enable Functional Parameter display
+
<b>Figure 1 </b>
−
===Functional Parameters===
+
 
−
<partinfo>BBa_K2350021 parameters</partinfo>
+
　　[[File:Holin-Endolysin-NrtA REcheck.png|200px]]
−
<!-- -->
+
 
 +
===Functional Test===
 +
　　As figure 2 shows, the trend of the relative absorbance is downward as the lactose is added to induce the suicide mechanism. The concentration of lactose is also positively correlated with the declining degree of relative absorbance (Experiment and graph provided by TAS_Taipei).
 +
 
 +
 
 +
<b>Figure 2 </b>
 +
 
 +
[[File:T--NYMU-Taipei--TAS-FunctionalTest.png|700px]]
 +
 
 +
 
 +
<partinfo>BBa_K2350021 SequenceAndFeatures</partinfo>

Latest revision as of 09:36, 1 November 2017


R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012-J23118-B0034-NrtA-B0015

Holin-Endolysin-NrtA

　　In this part, we combine holin (BBa_K2350019), endolysin (BBa_K2350020), and NrtA (BBa_K2350004). NrtA protein can stick to periplasmic membrane through a flexible linker to capture nitrite or nitrate in the periplasm. Therefore, we make use of NrtA in nitrogen starvation in our 2017 NYMU project. After we reach our goal, we need to kill E.coli to prevent contamination. When the lactose is added into the environment, the suicide mechanism, holin and endolyin, is induced. For more detail information, please see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, BBa_K2350019 for holin part, and BBa_K2350020 for endolysin-holin part.


Promoter, RBS, and Terminator

　　For more details of the connected promoter, RBS, and terminator, you can see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, and BBa_K2350019 for holin part.


Result

　　Figure 1 is restriction enzyme check electrophoresis result of Holin-Endolysin-NrtA construct. We use XmaI to digest sample 1~3. The result shows that 2 and 3 are right. M represents 1 kb marker.


Figure 1

　　Holin-Endolysin-NrtA REcheck.png

Functional Test

　　As figure 2 shows, the trend of the relative absorbance is downward as the lactose is added to induce the suicide mechanism. The concentration of lactose is also positively correlated with the declining degree of relative absorbance (Experiment and graph provided by TAS_Taipei).


Figure 2

T--NYMU-Taipei--TAS-FunctionalTest.png



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1035
    Illegal NheI site found at 1058
    Illegal NheI site found at 1757
    Illegal NheI site found at 1780
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2990
    Illegal BamHI site found at 2360
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1362
    Illegal AgeI site found at 1432
  • 1000
    COMPATIBLE WITH RFC[1000]