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. 2009 Sep 2;106(38):16174–16179. doi: 10.1073/pnas.0901455106

Fig. 1.

Fig. 1.

Overexpression of SpoIVFB and Pro-σK in E. coli. (A) Accumulation of SpoIVFB with or without N-terminal cytTM. E. coli cells bearing pZR209 to produce cytTM-SpoIVFB-FLAG2-His6 (lanes 1 and 3) or pZR260 to produce SpoIVFB-FLAG2-His6 (lane 2) were collected 2 h after IPTG induction. Extracts from equivalent cell amounts (based on the OD600 of the culture) (lanes 1 and 2) or 100-fold less extract (lane 3) were immunoblotted using antibodies against FLAG. (B) Cleavage of Pro-σK (1–126)-His6 by TM-SpoIVFB. E. coli cells bearing pZR327 to produce Pro-σK (1–126)-His6 S20G alone (lane 1) or in combination with pZR209 to also produce TM-SpoIVFB (lane 3), or bearing pZR12 to produce wild-type (WT) Pro-σK (1–126)-His6 in combination with pZR209 (lane 2), were collected 2 h after IPTG induction. Extracts from equivalent cell amounts were immunoblotted using antibodies against FLAG (top panel) or penta-His (bottom panel).