Part:BBa_K2765043

Produce redox sensitive Green fluorescent protein

Introduction

We obtained the DNA sequence of roGFP2+linker from PART: BBa_K2296006: Constitutive Promoter-RBS-roGFP2 - Orp1 C82S and codon optimized according to our chassis organism, Saccharomyces Cerevisiae, so that it can be better expressed in yeast.

At the same time, we introduced the gene sequences before and after optimization into yeast (the promoters are the same and the terminator are CYC1t). We selected four promoters of different intensities, adding enough DTT to the medium which made the proteins all in a reduced state. Then we measured the fluorescence intensity of roGFP2-Orp1 cited by at 488 nm (reduction peak) and the emission is at 515nm. Thus, the magnitude of the fluorescence intensity can reflect the level of protein expression to some extent.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 843
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1155