Part:BBa_K2971004

crtEBI under pBad promotor

This composite part contains the genes crtE, crtB and crtI combined into a single reading frame to produce lycopene when expressed in Escherichia coli. The genes express geranylgeranyl diphosphate synthase (GGPPS), phytoene synthase (CrtB) and phytoene desaturase (CrtI) respectively. GGPPS and CrtB produce the saturated compound phytoene, which is then desaturated by CrtI into the conjugated compound lycopene. When the part was expressed in E.coli, the pellet of the spun down expression culture acquired a pink color. The color validate that the part functions as intended.

Figure 1: Expression of pBAD-crtEBI in Escherichia coli (BL21)
Expression was induced in 2% arabinose
From left to right :Uninduced E. coli with empty expression vector, Induced E.coli with empty expression vector, induced E.coli with crtEBI expression vector, uninduced E.coli with crtEBI expression vector.

Figure 1: Colony PCR of cells transformed with crtEBI
Only one colony (8) showed the presence of the correct insert. 8 was then further investigated.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1205
  Illegal NheI site found at 4181
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2854
  Illegal BamHI site found at 1144
  Illegal BamHI site found at 2093
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1295
  Illegal SapI site found at 961