Part:BBa_K1933000

N-terminal domain of BclA protein fused to 6xHis tag

This part codes for N-terminal domain of BclA protein which is expressed on the cell surface connected by 6×His tag to be easily identified by Western blotting[1]. This domain does not have signal peptide for anchoring on the cell surface. Surface displayed passengers retain enzyme activity even when it is linked to BclA with 6× His tag.
For more information, please visit [http://2016.igem.org/Team:Kyoto our wiki].

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

BclA is a hair-like protein derived from Bacillus anthracis (34F2) that covers the B. anthracis’ spores. When BclA is surface expressed, their N-terminal domains (NTD) faces the inner cell, C-terminal domains (CTD) faces outwards, and internal repeating domains fill the gap between NTD and CTD.
A protein of interest can be displayed on the cell surface when it is linked to the C-terminus of N-terminal domain (21 amino acids) of BclA. Although most anchoring domains are limited in its ability to carry large passenger proteins, this domain is unique in that they can carry up to 120 kDa[1].
In our project, we used this domain to express cellulose binding domain (CBD) and anti-Norovirus scFv on the cell surface of E.coli.

We used this part for construction of BBa_K1933102, BBa_K1933103, and BBa_K1933201.

Characterization

Sequence confirmed!

Please see BBa_K1933102, BBa_K1933103 and BBa_K1933201 for full characterization.

Reference

[1] Park, Tae Jung, et al. "Surface display of recombinant proteins on Escherichia coli by BclA exosporium of Bacillus anthracis. " Microbial cell factories, 12.1, (2013): 1.