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Figure 4 | Microbial Cell Factories

Figure 4

From: Overproduction of Bacillus amyloliquefaciens extracellular glutamyl-endopeptidase as a result of ectopic multi-copy insertion of an efficiently-expressed mpr gene into the Bacillus subtilis chromosome

Figure 4

Southern hybridization of the chromosomal DNAs and SDS-PAGE of extracellular proteins secreted by the strains with different copy-number (N) of the integrated mprB.amy-cassettes. A – Hybridization with DNAs isolated from cells carrying (indicated as 1.; 2.; 3; etc. (N = 1: lane 1 (1.); N = 2: lane 2 (1.+2.); N = 4: lane 3 (1.+2.+3.+4.); and N = 7: lane 4 (1.+2.+3.+4.+5.+6.+7.) and lane 6 (1.+2.+3.+4.+5.+6.+8.). B SDS-PAGE of extracellular proteins secreted by the strains with: N = 0 (lane 2), N = 1 (lane 3), N = 2 (lane 4), N = 3 (lane 5), N = 9 (lane 8) and N = 10 (lane 9) mprB.amy cassettes in the chromosome or carrying the multi-copy-number recombinant plasmid pHE52mpr (lanes 6 and 10). Lanes 1 and 7-reference proteins with molecular mass given in kDa. The mature 220 amino acid B. amiloliquefaciens GSP has a molecular mass of about 23.7 kDa. The indicated extracellular GSP presented slightly lower electrophoretic mobility corresponding to 27-28 kDa; this was previously documented for the mature GSP from B. subtilis [17].

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