Part:BBa_K2986007

mRuby

Usage and Biology:

mRuby has the excitation and emission maxima at 558 nm and 605 nm, and a large Stokes shift of 47 nm, mRuby appears particularly useful for imaging applications.(Kredel, S., 2009). It is red fluorescent protein. This made the visualization of the target gene expression possible if we ligated the hGluc with target gene.

Design

We wanted to design a gene expression system with mRuby as reporter, so that we can simplify the production detection process into fluorescent observation. We designed a plasmid ligate our target gene with mRuby using mammalian lentivirus expression vector. We use a 45bp linker to connect the hGluc with the CD11b transmembrane domain and use P2A (2A peptide allows an open reading frame (ORF) to translate a peptide chain into several independent peptide chains) connect with Interleukin 8 CDS, in our assumption this allow the stable expression of both our target gene and the hGluc in Hela cells.

Location of features

Figure1.the plasmid used with hgluc

mRuby: 2379-3089 hGluc: 3120-3710
linker: 3711-3755
CD11b transmembrane domain: 3756-3989
P2A :3990-4055
Interleukin 8 CDS :4056-4352

Sequence and Features

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 16
  

References

Kredel, S., Oswald, F., Nienhaus, K., Deuschle, K., Röcker, C., Wolff, M., … Wiedenmann, J. (2009). mRuby, a bright monomeric red fluorescent protein for labeling of subcellular structures. PloS one, 4(2), e4391. doi:10.1371/journal.pone.0004391