Regulatory

Part:BBa_K5159006

Designed by: Tianyue WU   Group: iGEM24_HKUST   (2024-09-26)

Yarrowia lipolytica pIDP2 (YALI0F04095g) - Isocitrate Dehydrogenase Promoter

Endogenous promoter function: Drives the transcription of isocitrate dehydrogenase (IDP2), an enzyme involved in the TCA cycle that catalyzes the oxidative decarboxylation of isocitrate to alpha-ketoglutarate, producing NADPH.

Strength: Medium-strength promoter.

Activity Value: 1.05 x 10^7 RLU (relative light unit), which is the relative detection value of the amount of fluorescence light produced in each sample. The measurement was carried out with Nano-Glo® Luciferase Assay System kit, purchased from Promega (Catalog: #N1120).

Regulation: Displays increasing transcriptional activity during the exponential growth phase.

Role in Metabolism: Essential for the continuation of the TCA cycle, contributing to energy production and the generation of precursors for biosynthesis.

We chose 2 native inducible promoters, pCIT1 (Part:BBa_K3868074) and pIDP2, to be tested and potentially improved in our project, referencing to a research by Gu et al (2023). Both promoters are significant in cellular metabolism, particularly in the TCA cycle, and are subject to regulation under different environmental conditions, such as nitrogen depletion. The study provides insights into their potential use in biomanufacturing applications within the yeast Yarrowia lipolytica.

The main limitations of current studies on the Yarrowia lipolytica promoters are that the analysis is restricted to a short 1500 bp region upstream of the protein-coding gene, which may not encompass all necessary regulatory elements for full transcriptional control. The characterization of core promoter elements, such as the TATA box and initiator, remains unclear, making it challenging to understand the mechanisms governing transcription initiation.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 764
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1482
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 270
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 764
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 764
  • 1000
    COMPATIBLE WITH RFC[1000]

Reference

[1] Chang Wang, Mingxin Lin, Zhiliang Yang, Xueyao Lu, Yinfang Liu, Huizhi Lu, Jiang Zhu, Xiaoman Sun, Yang Gu (2023). Characterization of the endogenous promoters in Yarrowia lipolytica for the biomanufacturing applications. Process Biochemistry, Volume 124, 245-252. https://doi.org/10.1016/j.procbio.2022.11.023


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