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From 2010.igem.org

• Home
• Project
  • About The Toolkit
  • Oursystem
  • Producing The Antigen
  • Future Applications
  • Safety
• Toolkit
• Why Pichia
  • Pichia Vs Saccharomyces
  • Easy and Inexpensive to Culture
  • High production of foreign Protein while low levels of endogenous protein
  • Pichia pastoris versus Esherichia coli
• Protocols
  • Preparation Of Electrocompetent Cells
  • Preparation of Heat Shock Competent Cells
  • Transformation Using Electroporation
  • Transformation Using Heat Shock
• The Team
• Notebook

Designing the Antigen

In order to design an antigen we compared previously isolated and sequenced Influenza A H1N1 strains. We were able to identify a conserved region that comprised of the Hemagglutinin globular head which is formed by a disulfide bond between two cysteins. This region of the virus has been shown to produce an immune response. This is important since we plan on being able to use this as a potential vaccine. In order to identify the location of the cysteins within the sequence, we converted the nucleotide sequence to the amino acid sequence. The cysteins were identified, and our part was designed to include this region. The next step was to design this part to fit the registry standard. Two unwanted restriction sites were identified. In order to eliminate these sites, we did a nucleotide swap while maintaining the same amino acid sequence. Following the above steps, the part has been designed to be compatible with all assembly standards.