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. 2022 Feb 3;6:10. doi: 10.1038/s41538-022-00128-4

Fig. 1. Expression and identification of rsfAFP in B. subtilis WB800N.

Fig. 1

A PCR detects positive clones. M: DNA marker DL10000; lanes 1–7: recombinant plasmid; +: positive control. B Expression result. Coomassie blue staining. MW: molecular weight marker. ϕ: Transformed cells not induced. C Western blot detects expression results. MW: molecular weight marker. ϕ: transformed cells not induced. +: positive control. D Target protein purification profile. IN: input; FT: flow through; MW: molecular weight marker; W1– W3: wash with TBS buffer with 0, 30, and 50 mM imidazole, respectively; E1–E5: eluted with TBS buffer with 100, 150, 200, 250, and 400 mM imidazole, respectively. E Sequence cover situation of rsfAFP by Nano LC-MS/MS spectrometry. The red section indicates the peptides matching the peptides of the snow flea antifreeze protein. The gray section indicates the missing fragments in the Nano LC-MS/MS test. The green section of peptide is GS linker. The blue section of peptide is 6*his-tag.