Part:BBa_K1351006:Design

CWB domain of B. subtilis major autolysin LytC

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]

Design Notes

Due to the N-terminal signal peptide, this part allows only C-terminal fusions of proteins to be displayed on the cell surface.

The CWB domain of LytC consists of three N‐terminal type II cell wall‐binding motifs, ranging from position 30 to 318. To minimize possible effects of a translational fusion to the CWB domain, the part includes the downstream unstructered region until position 333. An additional linker can be inserted to allow more structural flexibilty.

Source

This part was generated by amplification from B. subtilis W168 gDNA with the primers listed below, followed by digestion with EcoRI and PstI and ligation into pSB1C3.

LytC_ENX_SD_Ngo_fwd: gatcGAATTCgcggccgctTCTAGAgtaaggaggaaGCCGGC TTGCGTTCTTATATAAAAGTCCTAACAATGTG

LytC_333_SNP_Age_rev: gatcCTGCAGcggccgctACTAGTattaACCGGT TGAATCTTGATCACCGTGACCC

References

Tsuchiya, Atsushi, Gota Kobayashi, Hiroki Yamamoto and Junichi Sekiguchi. "Production of a Recombinant Lipase Artificially Localized on the Bacillus Subtilis Cell Surface." FEMS Microbiology Letters 176, no. 2 (1999): 373-378.

Yamamoto, H., S. Kurosawa and J. Sekiguchi. "Localization of the Vegetative Cell Wall Hydrolases Lytc, Lyte, and Lytf on the Bacillus Subtilis Cell Surface and Stability of These Enzymes to Cell Wall-Bound or Extracellular Proteases." J Bacteriol 185, no. 22 (2003): 6666-77. [http://www.ncbi.nlm.nih.gov/pubmed/14594841 PubMed]