Regulatory

Part:BBa_K3804001

Designed by: Rintaro Shimojo   Group: iGEM21_UTokyo   (2021-09-25)


Yeast Anb1 promoter

There are many genes in yeast that are regulated by oxygens supply. ANB1 is one of the genes controlled by oxygen concentration. Many kinds of oxygen-dependent repressors are bind to the upstream region of the ANB1 ORF and regulate the expression. Especially, ROX1 repressor protein takes a major role in this regulation. This part is about 0.5kbp upstream region of ANB1 ORF and it works as a promoter. In the plasmid inserted environment, the Anb1 promoter functions as a promoter that is repressed under anaerobic conditions.

Characterization

As shown in Figure 1 and 2, this promoter responds to the change of oxygen concentration. The change of expression was indicated by mCherry FP. This part, pAnb1 promoter works sufficiently as can be seen in Figure1. And also, in this experiment, there is a possibility that the constitutive promoter(BBa_I766555) which we used for this experiment has oxygen-dependent activity change[4]. So further experiments are needed.

Figure1. The change of promoters activity under aerobic and anaerobic condition. pCYC:Constitutive promoter for control. pRox1:ROX1 upstreame region. pAnb1:ANB1 upstream region (This part).

Figure2. Change of activity of pCyc(BBa_I766555), pAnb1(BBa_K3804001) and pRox1(BBa_K3804000) indicated by fluorescence protein(mCherry) under aerobic and anaerobic condition. (40x,NA 0.95)

Reference

[1]Z.-A. Fang, G.-H. Wang, A.-L. Chen, Y.-F. Li, J.-P. Liu, and Y.-Y. Li, “Gene Responses to Oxygen Availability in Kluyveromyces lactis: an Insight on the Evolution of the Oxygen-Responding System in Yeast,” PLoS ONE, vol. 4, no. 10, p. 12, 2009
[2]C. V. Lowry, M. E. Cerdán, and R. S. Zitomer, “A hypoxic consensus operator and a constitutive activation region regulate the ANB1 gene of Saccharomyces cerevisiae,” Mol. Cell. Biol., vol. 10, no. 11, pp. 5921–5926, Nov. 1990, doi: 10.1128/mcb.10.11.5921-5926.1990.
[3]K. E. Kwast, P. V. Burke, and R. O. Poyton, “OXYGEN SENSING AND THE TRANSCRIPTIONAL REGULATION OF OXYGEN- RESPONSIVE GENES IN YEAST,” p. 19.
[4]U. Oechsner, H. Hermann, A. Zollner, A. Haid, and W. Bandlow, “Expression of yeast cytochrome cl is controlled at the transcriptional level by glucose, oxygen and haem,” p. 13.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 297
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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