Skip to main content
. 2011 May 26;6(5):e20327. doi: 10.1371/journal.pone.0020327

Figure 4. Expression and purification of OsSpo11 and OsTopVIB proteins.

Figure 4

Proteins were separated by 10% SDS-PAGE and stained with Coomassie Brilliant Blue (A∼C) or hybridized with an antibody against GST (D). A, Expression of the 4 proteins each in S. prombe cells (OsSpo11-4 showed here as an example). Cells harboring recombinant plasmids were treated with (+vb1) or without (−vb1) vitamin B1. GST-OsSpo11-4 fusion protein from −vb1 was purified with Glutathione Sepharose 4B resin. B, Removal of GST tag from purified GST-protein fusion by thrombin digestion (OsSpo11-4 shown as an example). The reaction mixture was incubated for different times shown above the image and subjected to SDS-PAGE separation. C, Purified GST-tagged OsSpo11s and OsTopVIB. D, Western blot analysis of purified GST-tagged proteins in C with a monoclonal antibody against GST. MW, molecular weight standards.