Part:BBa_K2543010

Ac5-hCD4-dToll-polyA / pSB1C3

- Ac5 is a strong and constitutive promoter from Drosophila actin 5c gene and commonly used in insect expression system. Human CD4 (hCD4) is a cell marker expressed on the subtype of T helper cell. CD4 acts as a coreceptor to help T cell development and cell function.
- CD4 plays an important role in T cell activation and immune signaling. The extracellular domain of hCD4 (1-396 aa) can form dimer and regulate the function of T cell activation.
- Toll is a transmembrane protein involved in insect immune defense system to recognize pathogens like bacteria, viruses and fungi. Toll activated by pathogens transmits the signaling to express anti-microbial peptide (AMP) to kill the pathogens. - - Drosophila transmembrane domain (808-828 aa) and intracellular domain (829-1097 aa) of Toll (dToll) play an important roll in regulating the immune signaling.
- Polyadenylation (polyA) is one of the process of eukaryotic mRNA translation. It adds a poly(A) tail to protect mRNA from enzymatic degradation and aid in transcription termination. Polyadenylation signal of simian virus 40 (SV40 poly A) has been used widely in mammalian and many eukaryotic gene expression system.
- This construct creates a synthetic human CD4-drosophila Toll chimera receptor system. The system functions not only in response to human HIV virus and also transmit Toll signaling to activate the expression of anti-microbial peptide (AMP)

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 104
  Illegal XhoI site found at 3691
  Illegal XhoI site found at 4153
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3760
  Illegal NgoMIV site found at 5079
  Illegal AgeI site found at 3742
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2923
  Illegal BsaI site found at 4549
  Illegal BsaI.rc site found at 2533
  

References

1. UniProtKB - P01730 (CD4_HUMAN)
2. Retrovirology. (2006) Association between disruption of CD4 receptor dimerization and increased human immunodeficiency virus type 1 entry
3. J Immunol. (2006) Evidence for a domain-swapped CD4 dimer as the coreceptor for binding to class II MHC.
4. J Immunol. (2006) Triggering of T cell activation via CD4 dimers.
5. J Biol Chem. (2014) Disulfide reduction in CD4 domain 1 or 2 is essential for interaction with HIV glycoprotein 120 (gp120), which impairs thioredoxin-driven CD4 dimerization.

1. UniProtKB - P08953 (TOLL_DROME)
2. Cell. (1988) The Toll gene of Drosophila, required for dorsal-ventral embryonic polarity, appears to encode a transmembrane protein.
3. Nat Rev Immunol. (2006) Toll-like receptors as molecular switches.
4. J Immunol. (2011) The Drosophila Toll signaling pathway.