Category:Team Phergalicious

Figure 2F shows a western blot performed after IPTG induction to detect the presence of HolFL by using anti-6xHis antibodies and Gp10 with anti-gp10 antibodies. pSKHol+gp10 shows higher expression between 42kDa and above, it also shows higher expression between 22kDa and 14kDa.

In Figure 4, the membrane fraction column (M) of the Western blot panel (bottom row) for gp 24.1 shows that the holin-like protein of Bacillus phage SPP1 was detected at much higher levels in the cell membrane after cell fractionation, as opposed to the cytoplasm. Cell fractionation into cytoplasmic and membrane fractions followed by Western blot shows a thicker band of protein in the membrane fraction of the cells expressing gp24.1, with no band visible in the cytoplasmic fraction.

Figure 3d shows the expression of HolGH15 over 10 minute intervals after western blotting was performed. The E. coli BL21-CodonPlus cells harbouring pET-28a-HOLGH15 were induced with IPTG to detect HolGH15. The result demonstrated the presence of a band of about 21 kDa with consistent size with the predicted mass of HolGH15. The band was detected 10 minutes after the induction with IPTG. Figure 3b: Transmission Electron Microscopy observations showed damage and disruption in the induced E. coli cells. The result was the release of cellular contents and showed apperance simillar to empty, non-living bacterial envelopes of Gram-negative bacteria.

In Figure 4, the membrane fraction column (M) of the Western blot panel (bottom row) for gp26 shows that the protein product was detected at much higher levels in the cell membrane, as opposed to the cytoplasm. Cell fractionation into cytoplasmic and membrane fractions followed by Western blot shows a thicker band of protein in the membrane fraction of the cells expressing gp26.