Part:BBa_K1728024

rhodopsin siganl peptide +IL-8 receptor type alpha (CXCR1)

rhodopsin siganl peptide +IL-8 receptor type alpha (CXCR1)

Usage and Biology

The rhodopsin-CXCR1 sequence is a combination of bovine rhodopsin signal peptide (BBa_K1728020) and CXCR1 with optimized sequence(BBa_K1728021)

Western blot analysis of Rho-CXCR1 expression

The galactose induction of CXCR1 expression was performed as described in protocol. We collect yeast samples at different induction time. Protein extracted from the yeast whole-cell extract was separated on SDS-PAGE and finally detected by anti-CXCR1 antibody.

Effect of IL-8 treatment in engineered yeast with Rho-CXCR1 expression

Plasmid p426GAL-CXCR1 was transformed into engineered yeast, and CXCR1 was induced by 2 % galactose overnight. At first, we could not see GFP production. In order to exclude the possibility that cell wall blocks the binding between CXCR1 and IL-8, yeast cells were digested with zymolyase to remove cell wall and became spheroplasts. After that, we add 1 μM IL-8 to stimulate GPCR-mediated pathway, we observed GFP after IL-8 treatment for 8 hours. The negative control is the same yeast strain without IL-8 treatment. Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 852
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 277
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 49
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1041