Part:BBa_K3888011

[Ni-Fe] Hydrogenase Core gene HoxG1 and HoxK1

This composite part is used for heterologous expression of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase in Escherichia coli.
Hydrogenase is extremely sensitive to oxygen. By expressing oxygen-tolerant hydrogenase in E.coli, the oxygen tolerance will increase thus increases the amount of hydrogen produced by hydrogenase in E.coli. The strain of E.coli we used in the experiments is E.coli BL21. The part basically consists of two main parts: HoxG1(BBa_K3888003) and HoxG1(BBa_K3888004), which encode the large subunit and small subunit of oxygen-tolerant Hydrogenovibrio marinus [NiFe]-hydrogenase respectively. Other key features also include a natural (wild) type of RBS, B0032m, and a pTac promoter which is controlled by LacI(induced by IPTG) and lac operator .

Figure 1: The Oxygen-tolerant Hydrogenase introduction plasmid constructed, which has a length of 8261bp.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 363
  Illegal XbaI site found at 465
  Illegal SpeI site found at 531
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 531
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 678
  Illegal BamHI site found at 1593
  Illegal BamHI site found at 2932
  Illegal XhoI site found at 1915
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 363
  Illegal XbaI site found at 465
  Illegal SpeI site found at 531
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 363
  Illegal XbaI site found at 465
  Illegal SpeI site found at 531
  Illegal AgeI site found at 1809
  
• 1000
  COMPATIBLE WITH RFC[1000]