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.

HB36.5 flu binder tested using Yeast surface display at 100 nM H2 on the left, H312A mutant tested on the right at 100 nM H2 on the right. HB36.5 Shows little to no binding at this HA concentration. H312A dramatically increases binding to H2.

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]