Part:BBa_K3629001

Yarrowia lipolytica TEF1 promoter with an intron

Promoter from the Yarrowia lipolytica Translation Elongation Factor 1 (TEF1) gene with the first intron.

Usage and Biology

This is the natural promoter found in the wild-type Yarrowia lipolytica for the Translation Elongation Factor 1 (TEF1) gene. TEF promoter is among the strongest constitutive promoters found in Yarrowia lipolytica. Inclusion of the first intron of the TEF1 gene in the promoter sequence has shown increase the expression levels of approximately 17 fold over the wild-type TEF1 promoter (BBa_K2117000) (2). It can be placed immediately upstream of a coding sequence for Y. lipolytica.

This promoter was used in the following expression constructs in our collection:

BBa_K3529012= T. reesei CBHII expression construct
BBa_K3529013= Modified P. funiculosum CBHI expression construct
BBa_K3529014= N. crassa CBHI expression construct
BBa_K3529016= Modified T. reesei EGI expression construct
BBa_K3529023= 2-isopropylmalate synthase (LEU4) overexpression construct
BBa_K3529024= Anthranilate synthase component 1 (TRP2) overexpression construct
BBa_K3529025= mCherry expression construct
BBa_K3529026= mCitrine expression construct

Previous studies incorporating cellulases in Y. lipolytica found this promoter to enhance the expression of cellulase enzymes, particularly CBHI (2).

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
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

The wild-type sequence of this promoter contains BsaI, PstI, and SpeI restriction sites in the promoter section before the intron sequence. Therefore, single-nucleotides changes that match the ones in the functional BBa_K2983050 promoter were made to remove these sites.

References

1. Blazeck, J., Liu, L., Redden, H., & Alper, H. (2011). Tuning gene expression in Yarrowia lipolytica by a hybrid promoter approach. Applied and environmental microbiology, 77(22), 7905–7914. https://doi.org/10.1128/AEM.05763-11

2. Wei H, Wang W, Alper HS, et al. Ameliorating the Metabolic Burden of the Co-expression of Secreted Fungal Cellulases in a High Lipid-Accumulating Yarrowia lipolytica Strain by Medium C/N Ratio and a Chemical Chaperone. Front Microbiol. 2019;9:3276. Published 2019 Jan 9. doi:10.3389/fmicb.2018.03276