Part:BBa_K3165006

ECFP (Optimized for Escherichia coli)

ECFP or Engineered Cyan Fluorescent Protein is a monomeric protein artificially derived from the naturally occurring Green Fluorescent Protein (GFP) in the chemiluminescent bacteria, Aequorea victoria. Developed by the specific mutagenesis of the GFP gene, ECFP has increased stability to varying temperatures, sustained fluorescence and relatively shorter maturation times which makes it a suitable fluorophore.

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]

Biology

Green Fluorescent Protein derived from the chemiluminescnet bacteria Aequorea victoria, is one of the most widely used fluorophores in molecular biology and biochemistry experiments. Since the discovery of GFP, several variants of the protein have been developed to suit particular requirements of different experiments. ECFP, a monomeric protein belongs to the Cyan Fluorescnet Protein family (where Tyr-66 is replace dwith a Trp). Due to its increased tolerance over a wide range of temperature, stability to photobleaching and relatively shorter maturation time, it is commonly used a fluorescent marker for biological studies. ECFP also forms a popular FRET pair along with EYFP (engineered yellow fluorescent protein).

Usage

Fluorescent proteins like ECFP are used as non-invasive markers in various cell lineages to study gene expression, protein-protein interactions and signalling & secretion mechanisms. This part can be used as a reporter protein to characterise the optogenetic system incorporated in our model organism. Fluorescence caused by ECFP can be used as parameter to quantify the activity of the photo-sensitive system (CcaSR in E. coli).

References :

(1) https://www.fpbase.org/protein/ecfp/
(2) Heim, R., & Tsien, R. Y. (1996). Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer. Current Biology, 6(2), 178–182. doi:10.1016/s0960-9822(02)00450-5 
(3) Tsien, R. Y. (1998). THE GREEN FLUORESCENT PROTEIN. Annual Review of Biochemistry, 67(1), 509–544. doi:10.1146/annurev.biochem.67.1.509