Difference between revisions of "Part:BBa K3804000"
 
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<partinfo>BBa_K3804000 short</partinfo>
 
<partinfo>BBa_K3804000 short</partinfo>
  
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There are many genes in yeast that are regulated by oxygens supply. ROX1 is one of the most important factors in the process of oxygen response and it works as the repressor under the aerobic condition. ROX1`s expression itself is also regulated by much regulatory protein and the upstream region of ROX1 functions as an oxygen-dependent promoter.
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There are many genes in yeast that are regulated by oxygen supply. ROX1 is one of the most important factors in the process of oxygen response and it works as the repressor under the aerobic condition. ROX1`s expression itself is also regulated by much regulatory protein and the upstream region of ROX1 functions as an oxygen-dependent promoter.
 
This part is about 1.3kbp upstream region of Rox1 ORF and it works as a promoter.  
 
This part is about 1.3kbp upstream region of Rox1 ORF and it works as a promoter.  
 
In the plasmid insertion environment, the Rox1 promoter functions as a promoter that is repressed under anaerobic conditions.
 
In the plasmid insertion environment, the Rox1 promoter functions as a promoter that is repressed under anaerobic conditions.
  
 
===Characterization===
 
===Characterization===
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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. However, the change of expression in pRox1(This part)is relatively small and it needs additional experiments to actual usage. And also, in this experiment, there is a possibility that the constitutive promoter which we used for this experiment has oxygen-dependent activity change[4]. So further experiments are needed.
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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. However, the change of expression in pRox1(This part)is relatively small and it needs additional experiments to actual usage. And also, in this experiment, there is a possibility that the constitutive promoter(<partinfo>BBa_I766555</partinfo>) which we used for this experiment has oxygen-dependent activity change[4]. So further experiments are needed.
  
 
[[File:T--UTokyo--OXYGraph.png|550px|thumb|center|Figure1. The change of promoters activity under aerobic and anaerobic condition. pCYC:Constitutive promoter for control. pRox1:ROX1 upstreame region(This part). pAnb1:ANB1 upstream region.]] <br>
 
[[File:T--UTokyo--OXYGraph.png|550px|thumb|center|Figure1. The change of promoters activity under aerobic and anaerobic condition. pCYC:Constitutive promoter for control. pRox1:ROX1 upstreame region(This part). pAnb1:ANB1 upstream region.]] <br>
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[[File:T--UTokyo--OXYFP.png|550px|thumb|center|Figure2. Change of activity of pRox1 indicated by fluorescence protein(mCherry) under aerobic and anaerobic condition. (40x,NA 0.95) ]]<br>
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[[File:T--UTokyo--OXYFP.png|550px|thumb|center|Figure2. Change of activity of pCyc(<partinfo>BBa_I766555</partinfo>), pAnb1(<partinfo>BBa_K3804001</partinfo>) and pRox1(<partinfo>BBa_K3804000</partinfo>) indicated by fluorescence protein(mCherry) under aerobic and anaerobic condition. (40x,NA 0.95) ]]<br>
  
 
===Reference===
 
===Reference===

Latest revision as of 14:06, 21 October 2021


Yeast Rox1 promoter

There are many genes in yeast that are regulated by oxygen supply. ROX1 is one of the most important factors in the process of oxygen response and it works as the repressor under the aerobic condition. ROX1`s expression itself is also regulated by much regulatory protein and the upstream region of ROX1 functions as an oxygen-dependent promoter. This part is about 1.3kbp upstream region of Rox1 ORF and it works as a promoter. In the plasmid insertion environment, the Rox1 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. However, the change of expression in pRox1(This part)is relatively small and it needs additional experiments to actual usage. 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(This part). pAnb1:ANB1 upstream region.

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]J. Deckert, A. M. Rodriguez Torres, J. T. Simon, and R. S. Zitomer, “Mutational analysis of Rox1, a DNA-bending repressor of hypoxic genes in Saccharomyces cerevisiae.,” Mol. Cell. Biol., vol. 15, no. 11, pp. 6109–6117, Nov. 1995.
[2]J. Deckert, R. Perini, B. Balasubramanian, and R. S. Zitomer, “Multiple elements and auto-repression regulate Rox1, a repressor of hypoxic genes in Saccharomyces cerevisiae.,” Genetics, vol. 139, no. 3, pp. 1149–1158, Mar. 1995, doi: 10.1093/genetics/139.3.1149.
[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
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]