Difference between revisions of "Part:BBa K1342001"
(→‎TLC Assay)
 
(4 intermediate revisions by the same user not shown)
Line 9: Line 9:
  
  
−
ZinTp_AspA contains ZinTp([https://parts.igem.org/Part:BBa_K1342005 BBa_K1342005]), RBS([https://parts.igem.org/Part:BBa_B0034 BBa_B0034]), ApeA([https://parts.igem.org/Part:BBa_K1342002 BBa_K1342002]), Dter([https://parts.igem.org/Part:BBa_B0015 BBa_B0015]).
+
ZinTp_AspA contains ZinTp([https://parts.igem.org/Part:BBa_K1342005 BBa_K1342005]), RBS([https://parts.igem.org/Part:BBa_B0034 BBa_B0034]), ApeA([https://parts.igem.org/Part:BBa_K1342002 BBa_K1342002]) and Dter([https://parts.igem.org/Part:BBa_B0015 BBa_B0015]).
−
AspA is expressed in the presence of cadmium ion. And translated to aspartase.
+
The AspA encoding aspartase that synthesize aspartic acid from fumaric acid and ammonium, is expressed in the presence of cadmium ion.  
  
  
Line 32: Line 32:
 
===TLC Assay===
 
===TLC Assay===
  
−
[[File:TLC_20140929_.jpg|200px|thumb|left|
+
 
−
fig.1
+
[[File:TLC_20140929_.jpg|200px|thumb|fig.1
 
<br>
 
<br>
−
Analysis of aspartate from strain E.coli K-12/[https://parts.igem.org/Part:BBa_K1342001 BBa_K1342001] by thin-layer chromatography (TLC) Lane: 1, not addition cadmium to previous culture of synthesis medium (diluted 1:50 by synthesis medium) 2, (diluted 1:100 by synthesis medium) 3, add cadmium (250μM) 4, (diluted 1:50 by synthesis medium) 5, (diluted 1:100 by synthesis medium) 6, (diluted 1:200 by synthesis medium)]]
+
Analysis of aspartic acid synthesized by <i>E. coli</i> JM109/BBa_K1342001 by thin-layer chromatography (TLC) Lane: 1, 2 no addition into culture (diluted 1:50 and 1:100 by synthesis medium, respectively) 3, additon of cadmium ion (250μM) 4, 5, 6 (diluted 1:50, 1:100 and 1:200 by synthesis medium, respectively)|left]]
  
  
Line 41: Line 41:
 
fig.2
 
fig.2
 
<br>
 
<br>
−
intensity of the band which is the Quantitative data of the aspartic acid synthesis.]]
+
Estimation of aspartic acid content in spot
 +
Each intensity of spots indicating the content of aspartic acid, was estimated by Image J.
 +
.]]

Latest revision as of 12:53, 17 October 2014

zinTp_AspA


Asp Device image.jpg


ZinTp_AspA contains ZinTp(BBa_K1342005), RBS(BBa_B0034), ApeA(BBa_K1342002) and Dter(BBa_B0015). The AspA encoding aspartase that synthesize aspartic acid from fumaric acid and ammonium, is expressed in the presence of cadmium ion.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 179
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1071
    Illegal AgeI site found at 600
    Illegal AgeI site found at 1452
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1111



TLC Assay

fig.1
Analysis of aspartic acid synthesized by E. coli JM109/BBa_K1342001 by thin-layer chromatography (TLC) Lane: 1, 2 no addition into culture (diluted 1:50 and 1:100 by synthesis medium, respectively) 3, additon of cadmium ion (250μM) 4, 5, 6 (diluted 1:50, 1:100 and 1:200 by synthesis medium, respectively)


fig.2
Estimation of aspartic acid content in spot Each intensity of spots indicating the content of aspartic acid, was estimated by Image J. .