Part:BBa_K2257000

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.

Design

This design is Inspired from Cupriavidusnecator H16.

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

File:T-Nanjing-China-h2-Data 1.tif

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