Part:BBa_P0140

PoPS -> TetR [S0163]

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]

Tianjin's 2021 characterization

New information of TetR operon leakage

Group: Tianjin 2021
Author: Ruiqi Liu, Lingwei Ding
Summary: we added information about TetR operon leakage.

Background

Figure 1.Carotene plasmid with TetR operon

In our project, we want to use TetR operon to control the expression of mcherry gene. Thus, we want to test whether TetR operon can strictly regulate the expression of gene or not. We constructed a carotene plasmid with TetR operon, and transformed it into Saccharomyces cerevisiae. If the carotene gene controlled by TetR operon leaks, the yeasts will turn orange.

Result

We cultured the yeasts after transforming the carotene plasmid. We were surprised to find that, before using Atc inducer, a large proportion of yeasts was orange. Our experimental results show that the tetR promoter is very easy to leak, and the expression amount of the circuit is different and difficult to control. Therefore, the other iGEMers should aware that TetR operon is not a good choice if you want to strictly control the expression of structure gene.

Figure 2.Saccharomyces cerevisiae with carotene plasmids shows that TetR operon is easy to leak