Part:BBa_K892403

HB36.5_H312A

A mutated HB80.4 flu binder aimed to increase binding to H2, a subtype of hemagglutinin on the surface of the Influenza virus. This mutant has a point mutation at residue 312 from Histidine to Alanine, which dramatically increases the binding to H2 over the native HB36.5 binder.

Usage and Biology

This part was tested by the [http://2012.igem.org/Team:Washington 2012 University of Washington] iGEM team to bind to various subtypes of Hemagglutinin in Group 1.To test BBa _K892401, it was inserted into a protein expression vector, pETCON. HB36.5_H312A was then produced and transformed into yeast as described in the [http://2012.igem.org/Team:Washington/Protocols/Yeast 2012 Team's Yeast Transformation Protocol]. The binding protein was then tested for activity against H2. For a detailed description of the methodology, please see the [http://2012.igem.org/Team:Washington/Protocols/Display 2012 UW iGEM Yeast Surface Display protocol]. The resulting data is shown below.

HB36.5 and its variants tested against hemagglutinin subtype 2. HB36.5 is shown to have very little activity in binding H2. The single-point mutant H312A dramatically increased binding to H2 at all concentrations of HA tested.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 107
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 107
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 289
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 107
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 107
  
• 1000
  COMPATIBLE WITH RFC[1000]