SDS-PAGE analysis of proteins distributed between soluble and insoluble cell fraction after bacterial cell disruption (G-CSF). Target protein (G-CSF) was overexpressed in the form of ncIBs inside bacterial cells. Three mechanisms of bacterial cell disruption were studied: enzymatic lysis (L), high pressure homogenization (H) and sonication (S). After bacterial cell disruption soluble cell fraction (sn) and insoluble cell fraction (p) were separated with centrifugation and both samples were analysed with SDS-PAGE. Whole bacterial cells were also analysed (total cell proteins - Tcp). As standard (St) mixture of four proteins with known concentrations and known molecular weight was used. The arrow shows protein G-CSF. In the samples lysed with enzymatic lysis, the most abundant protein is enzyme Lysozyme. Legend: standard mixture (St) composition: BSA - Bovine serum albumin, 66.2 kDa (0.25 μg/gel). CA I - Carbonic anhydrase I, 31 kDa (0.5 μg/gel). M - Myoglobin, 17.2 kDa (1 μg/gel). L - Lysozyme, 14.4 kDa (1.5 μg/gel).
Peternel and Komel Microbial Cell Factories 2010 9:66 doi:10.1186/1475-2859-9-66