Difference between revisions of "Part:BBa K2257000"
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  <p><font size="+1">Now  we&rsquo;ve succefully detected the  protein expression by SDS-Page analysis and Western blot analysis.</font></p>
 
  <p><font size="+1">Now  we&rsquo;ve succefully detected the  protein expression by SDS-Page analysis and Western blot analysis.</font></p>
 
[[File:T-Nanjing-China-h2-7.png|460px|thumb|left|Figure 1. Coomassie  Brilliant Blue R-250-stained SDS-Page analysis of recombinant E.coli expressing  hoxABCJ-terminator-hoxp-gfp]]
 
[[File:T-Nanjing-China-h2-7.png|460px|thumb|left|Figure 1. Coomassie  Brilliant Blue R-250-stained SDS-Page analysis of recombinant E.coli expressing  hoxABCJ-terminator-hoxp-gfp]]
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[[File:T-Nanjing-China-h2-8.png|390px|thumb|right|Fingure 2. Western blot analysis of recombinant E.coli expressing his-hoxA]]</div>
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[[File:T-Nanjing-China-h2-8.png|380px|thumb|right|Fingure 2. Western blot analysis of recombinant E.coli expressing his-hoxA]]</div>
 
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Revision as of 15:03, 26 October 2017


hydrogen sensor sequence, HoxA-HoxB-HoxC-HoxJ-terminator-HoxP-EGFP

a composite of hydrogen sensor full length sequence. The order of the elements is: HoxA-HoxB-HoxC-HoxJ-terminator-HoxP-EGFP. The sequence of HoxABCJP comes from Ralstonia eutropha H16 megaplasmid pHG1. The hole sequence acts as an hydrogen sensor.

Usage and Biology

Design

This design is Inspired from Cupriavidusnecator H16.

T-Nanjing-China-h2-2.png

Experiments

Now we’ve succefully detected the protein expression by SDS-Page analysis and Western blot analysis.

Figure 1. Coomassie Brilliant Blue R-250-stained SDS-Page analysis of recombinant E.coli expressing hoxABCJ-terminator-hoxp-gfp
Fingure 2. Western blot analysis of recombinant E.coli expressing his-hoxA

Fluorescence intensity remains stationary when IPTG is added.
And Fluorescence intensity increases in a low hydrogen atmosphere.
When the amount of hydrogen goes to a higher level Fluorescence intensity increases apparently. meaning the designed reporter pathway have worked.

Figure 3. Influence of H2 concentration on fluorescence expression

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 5912
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 6989
    Illegal BamHI site found at 1
    Illegal XhoI site found at 409
    Illegal XhoI site found at 2562
    Illegal XhoI site found at 2574
    Illegal XhoI site found at 3102
    Illegal XhoI site found at 3129
    Illegal XhoI site found at 3876
    Illegal XhoI site found at 4152
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 816
    Illegal NgoMIV site found at 1767
    Illegal NgoMIV site found at 1959
    Illegal NgoMIV site found at 2074
    Illegal NgoMIV site found at 2343
    Illegal NgoMIV site found at 2751
    Illegal NgoMIV site found at 3501
    Illegal AgeI site found at 891
    Illegal AgeI site found at 1476
    Illegal AgeI site found at 3519
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 150
    Illegal BsaI.rc site found at 3199
    Illegal BsaI.rc site found at 3516
    Illegal BsaI.rc site found at 4347
    Illegal SapI.rc site found at 2304
    Illegal SapI.rc site found at 4991