Difference between revisions of "Part:BBa K1342002"
(→‎TLC Assay)
Line 27: Line 27:
 
===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 36: Line 36:
 
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 using Image J.
 +
.]]

Revision as of 13:20, 16 October 2014

AspA LVA(-)

Asp PDB.jpg
We removed LVA tag that renders protein susceptible to the action of tail-specific proteases from BBa_C0083 .
AspA is coding sequence of aspartase carries out reversible conversion of L-aspartate to fumarate and ammonia.


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 72
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 964
    Illegal AgeI site found at 493
    Illegal AgeI site found at 1345
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1004



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 using Image J. .