Part:BBa_K3552002

PaPilA

PaPilA is a type 4 pilus generally on Pseudomonas aeruginosa which can conduct electricity. This part is in the part collection where we provide different conductive pilus based on the same pilin generator. By these different type 4 pilus we can compare the qualities.

The part collection includes: Parts that are different kinds of type 4 pilus: BBa_K3552003 BBa_K3552004 BBa_K3552005 BBa_K3552006 BBa_K3552007 BBa_K3552008 BBa_K3552018 BBa_K3552019 BBa_K3552020 BBa_K3552021 BBa_K3552022 BBa_K3552023 BBa_K3552024 BBa_K3552025 BBa_K3552026 BBa_K3552027 BBa_K3552028 BBa_K3552029

Our part collection can instruct other teams to designed new rechargeable pilus and substitution of different major pilin.

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]

Usage and Biology

PaPilA are fimbrial protein which normally grow on Pseudomonas aeruginosa, but our PapilA is an artificially modified gene to enable it to conduct electricity. They are type 4 pili which are long and thin that displayed on the cell surface membrane. They can promote adherence, motility and transport functions in the bacteria. They are mainly built as helical polymers of a single subunit called the major pilin. These pilins are initially in the plasma membrane and the N-terminal is positively charged. The mature pilins will be extracted through the membrane by the pilin generator. The naturally occurred PaPilA contains ß-sheet and ∂ß-loop on the C-terminal which prevents them from conducting electricity at the beginning. This part is removed in our design to maintain same structure as GsPilA.

Characterization

We first constructed the PaPilA by SOE PCR through 8 different primers to maintain the exactly same sequence as in the Geobacter sulfurreducens. After the final construction of the generator of PilA, we added six His-tag on the PaPilA and expressed the gene in vivo. After 48 hours of growth, we extracted the pili and verified the existence of GsPilA on the outer cell through Western Blot. We also do electronic speculum and see the clear image of the pili in the solution.