Part:BBa_K4863004

PpsbA2_PilA1_SpyTag

A PilA1-SpyTag complex with SpyTag(BBa_K1159200) fused to the C-terminus of PilA1(BBa_K4863001) is constructed to achieve surface display of SpyTag on the cell surface of Synechocystis PCC 6803.

Usage and Biology

Fusion of a recombinant protein to the C-terminus of PilA1 will yield a cell covered with the fusion protein on its Type IV pili. Only small affinity proteins (<6.5kDA) can be functionally displayed via this system, or else the fusion protein would interfere with the biological functions of the pili. We present a system where SpyTag is modified to display on the cell surface of Synechocystis via fusion with PilA1.

PilA1 is a major protein subunit in Type IV pili (T4P), which is a surface-exposed appendage responsible for various essential functions in cyanobacteria,formed by polymerization of several pilin monomers. PilA1 constitutes the major pilin, which is the major structural component of T4P necessary for its biogenesis. SpyTag is a 13 amino acid-long peptide chain originally isolated from Streptococcus pyogenes.

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]