Part:BBa_K200011

Temperature Sensitive Lamda cI Repressor

The temperature sensitive lamda cI repressor protein has a cI857 mutation that results in denaturation of the repressor when the temperature is raised from 30 to 42°C, thereby allowing lambda promoter expression. cI1

The repressor normally negatively regulates the expression of genes from the bacteriophage lambda pL and pR promoters. This repressive action is strongest at 30°C. cI2

However, when the temperature is raised, typically to 42°C, the functionality of the protein is lost and the cI repressor is no longer able to bind to the operators on its promoter. Therefore, lambda promoter expression increases.

Measure management performed by iGEM22_HNU_China

We are team iGEM22_HNU_China. In the 2022 iGEM competition, we used the temperature sensing element cI857, that regulate an R promoter.

Here are our experimental introduction and results. You can also see our new part at BBa_K4252025.

Usage and Biology

The repressor is a single-chain, acidic protein containing 236 amino acids (Monomer Mr 26,000).

The N-terminal domain is responsible for DNA binding (operator site) and contact with RNA polymerase, while the C-terminal domain is responsible for dimer formation and higher order protein–protein contacts.

Requirements/Limitation

• More than one temperature sensitive mutations could lead to loss of DNA binding ability cI2

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]

References

<biblio> cI1 pmid=5274442 </biblio> <biblio> cI2 pmid=10235623 </biblio>