Part:BBa_K2022011

GFP, with Periplasm-localisation Tag

This part fuses a sec-signal tag from the pelB leader sequence (BBa_J32015) to GFP's n-terminus. This targets the protein to the sec pathway (Low et al., 2013) which helps export the fusion product to the periplasm and thus bias its subcellular localisation there.

Usage and Biology

Expression of this part will preferentially fill the periplasm with green fluorescence

Sequence and Feature Information

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 54
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 710
  

This part is simply an ORF containing GFP fused to an n-terminal sec signal peptide. To express it, it will need to be put into an expression vector or assembled with a promoter, RBS, and terminator.

Characterisation by UNSW Australia (2016)

Coming soon!

References

Lei, S.P., Lin, H.C., Wang, S.S., Callaway, J. and Wilcox, G., 1987. Characterization of the Erwinia carotovora pelB gene and its product pectate lyase. Journal of bacteriology, 169(9), pp.4379-4383.

Low, K.O., Mahadi, N.M. and Illias, R.M., 2013. Optimisation of signal peptide for recombinant protein secretion in bacterial hosts. Applied Microbiology and Biotechnology, 97(9), pp.3811-3826.