Difference between revisions of "Part:BBa K1342002"
 
(10 intermediate revisions by the same user not shown)
Line 4: Line 4:
 
[[File:Asp_PDB.jpg]]
 
[[File:Asp_PDB.jpg]]
 
<br>
 
<br>
−
We removed LVA tag that renders protein susceptible to the action of tail-specific proteases from [https://parts.igem.org/Part:BBa_C0083 BBa_C0083] .
+
 
 +
[https://parts.igem.org/Part:BBa_C0083 BBa_C0083] encodes aspartase with LVA tag, that tag renders protein susceptible to the action of tail-specific proteases.
 +
We removed LVA tag in [https://parts.igem.org/Part:BBa_C0083 BBa_C0083].
 
<br>
 
<br>
 
AspA is coding sequence of aspartase carries out reversible conversion of L-aspartate to fumarate and ammonia.
 
AspA is coding sequence of aspartase carries out reversible conversion of L-aspartate to fumarate and ammonia.
 +
 +
 
<br>
 
<br>
  
Line 25: Line 29:
  
 
<br>
 
<br>
−
[[File:TLC_20140929 .jpg]]
+
===TLC Assay===
 +
 
 +
 
 +
[[File:TLC_20140929_.jpg|200px|thumb|fig.1
 +
<br>
 +
Analysis of aspartic acid synthesized by <i>E. coli</i> JM109/[https://parts.igem.org/wiki/index.php?title=Part:BBa_K1342001 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]]
 +
 
 +
 
 +
 
 +
 
 +
[[File:TLC_intensity_of_the_band.jpg|400px|thumb|center|
 +
fig.2
 +
<br>
 +
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 20:53, 17 October 2014

AspA LVA(-)

Asp PDB.jpg

BBa_C0083 encodes aspartase with LVA tag, that tag renders protein susceptible to the action of tail-specific proteases. We removed LVA tag in 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 Image J. .