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Fig 4 | Microbial Cell Factories

Fig 4

From: Development of Pgrac100-based expression vectors allowing high protein production levels in Bacillus subtilis and relatively low basal expression in Escherichia coli

Fig 4

Overexpression and affinity purification of proteins fused to a His- or Strep-tag. B. subtilis 1012 carrying (a) pHT254-bgaB (BgaB-His-tag), (b) pHT254-gfp (GFP-His-tag), (c) pHT253-gfp (His-tag-GFP) and (d) pHT255-gfp (GFP-Strep-tag) were grown in LB medium to mid-log phase, and production of the recombinant proteins was induced by the addition of 0.1 mM IPTG. Cells were lysed, and aliquots were analyzed by SDS-PAGE (lane T, total cellular protein). The cellular extracts were applied to appropriate affinity columns, washed extensively and the bound protein was eluted as described under Materials and Methods. E1, E2 and E3 indicate the first, the second and the third elution step, respectively

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