Difference between revisions of "Part:BBa K1342002"

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[[File:Asp_PDB.jpg]]
 
[[File:Asp_PDB.jpg]]
 
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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] .
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[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.
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We removed LVA tag in [https://parts.igem.org/Part:BBa_C0083 BBa_C0083].
 
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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.
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[[File:TLC_20140929_.jpg|200px|thumb|fig.1
 
[[File:TLC_20140929_.jpg|200px|thumb|fig.1
 
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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]]
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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]]
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  Estimation of aspartic acid content in spot
 
  Estimation of aspartic acid content in spot
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Each intensity of spots indicating the content of aspartic acid, was estimated by using Image J.
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Each intensity of spots indicating the content of aspartic acid, was estimated by Image J.
 
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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. .