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Bacterial Expression System> | n. To obtain a plasmid that codes for full-length bactofilin protein from P. infestans (PiBac), the gene PITG_07992 (UniProtKB, D0N980) was synthesized (GenScript), amplified and cloned into the pHis17 plasmid using Gibson assembly (New England Biolabs), with a stop codon before the C-terminal hexahistidine tag on the plasmid. C41(DE3) E. coli cells (Lucigen) were transformed with the resulting plasmid by electroporation. | Get A Quote |
Bactofilins are small β-helical proteins that form cytoskeletal filaments in a range of bacteria. Bactofilins have diverse functions, from cell stalk formation in Caulobacter crescentus to chromosome segregation and motility in Myxococcus xanthus. However, the precise molecular architecture of bactofilin filaments has remained unclear. Here, sequence analysis and electron microscopy results reveal that, in addition to being widely distributed across bacteria and archaea, bactofilins are also present in a few eukaryotic lineages such as the Oomycetes. Electron cryomicroscopy analysis demonstrated that the sole bactofilin from Thermus thermophilus (TtBac) forms constitutive filaments that polymerize th... More