Part:BBa_K2520020

CMV-Mono Display:peanut 3-hGH

This device is a complete system for displaying proteins on cells' membrane. The Igk leader (BBa_K2520029) is a short signal peptide that prompts the translocation of a protein to the cellular membrane. PDGFR (BBa_K2520037) is a trans-membrane domain that anchors all the components located between the igk leader and the PDGFR itself to the membrane. His (BBa_K2520032) is a tag that binds to specific antibodies and provide an indirect method for verifying the display of proteins on the membrane. The protein that we chose to express on the membrane is an epitope that is known target of peanut allergy (BBa_K2520042, Arah2/6 protein) and is component related to our specific project. The CMV promoter (BBa_K1119006) is a constitutive expression promoter in mammalian cells and hGH (BBa_K404108) serves as a terminator.

Peanut Allergy

Allergies to peanuts are a major public health concern and life threatening. 1-2% of the population suffers from this allergy. Common symptoms include acute allergic rash, acute vomiting, laryngeal oedema, hypotension, and dysrhythmia. The major allergy inducing proteins in peanuts are generally considered to be Ara h 1 and Ara h 3 that are members of the cupin superfamily of proteins.

Epitopes

The major peanut allergens are Ara h1, Ara h2, and Ara h3

Peanut 3 (Ara h3):

Belongs to the legumin (11S) family and recognized by 50% of peanut-allergic. A hexamer (360–380 kD) formed by a head-to-head association of two trimers. Each monomer was found to have 4 linear epitopes.

Ara h 3 is represented with each of the monomers shown in a different color. In the gray monomer, linear epitope 1, 2, and 3 are shown in red, green, and blue, respectively

References

(1) Al-Muhsen, Saleh, Ann E. Clarke, and Rhoda S. Kagan. "Peanut allergy: an overview." Canadian Medical Association Journal 168.10 (2003): 1279-1285.

(2) Mueller, Geoffrey A., Soheila J. Maleki, and Lars C. Pedersen. "The molecular basis of peanut allergy." Current allergy and asthma reports 14.5 (2014): 1-9.‏

(3) Zhou, Yang, et al. "Peanut allergy, allergen composition, and methods of reducing allergenicity: A review." International journal of food science 2013 (2013).‏

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 614
  Illegal BamHI site found at 730
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 679
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1372