Part:BBa_K1072017

pGAL1 + Flag + odr-10 + GFP + ADH1Te + pFUS1 + BFP + ADH1Te

BBa_K1072017 is a composite device, including Odr-10 ,GFP and reporter gene, BFP.

Transforming Odr-10 into yeast and expressing,it active downstream pathway which is composed of Gpa1p (Gα), Ste4p (Gβ) and Ste18p (Gγ) subunits. The GPCR induces the complex of Gα and Gβγ, resulting in the separation of Gα and Gβγ. The Gβγ particle subsequently transmits the signal to a MAP kinase pathway, comprising Ste11p, Ste7p, and Fus3p held in a complex with the Ste5 protein. a reporter of this pathway was under the control of a Ste12p inducible promoter, Fus1 and link BFP.

Figure 2. Comparison between endogenous GPCR pathway in yeast and motified Odr10 pathway

After transforming the BBa_K1072017 into yeast.Fluorescence was detected.

Figure 5: Transforming Odr-10 with flag tag and reporter gene into yeast. GFP(a1,a2), flag tag signal(b1,b2) and BFP(c1,c2) was detected successfully, meaning that the whole pathway was stimulated by diacetyl.

This part is an improvement of BBa_K211002 and BBa_K775003 made by 2009 Hongkong university and 2012 TU delft team, respectively. A flag tag, GFP and report gene with promotor FUS1 have been added.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 150
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2258