Category:Team Red-A

The protein being tested AmyQ shows a lower concentration in the cell with the prsA3 mutation then compared to the wild type. Figure 2.

Figure 5:

It was previously proven, in the same paper, that orf9 is a N-acetylmuramoyl-L-alanine amidase, but it wasn't known what regions of orf9 was imporant for the lytic activity. To determine the regions of orf9 that are functionally important the researchers constructed eight mutants of the orf9 (of E. faecalis phage φEF24C) by deleting the N-terminal and C-terminal domains (Figure 1A). Only three of the C-terminal deleted mutants were viable (they were soluble), but none of the N-terminal regions were soluble which would indicate that the N-terminal region is required for stability. The C-terminal mutants had their lytic activities were tested by measuring the turbidity, but only the full length orf9, with a Histidine tag, displayed lytic activity. This demonstrated that the full length, N-terminal and C-terminal, orf9 is required for solubilty and lytic activity.

After finding a gene product termed yjbR they noticed that it had a sequence similarity with bacteria sarcosine oxidases. From there they measured yjbR gene products substrate specificity (table 2). They found that there is higher rate of activites with N-methyl-d-alanine, d-proline, d-alanine, and d-2-aminobutyrate.

In Figure 1C there is a graph that shows the amount of sporulation over time. The wild type that has a normal Skf gene shows a low amount of sporulation compared to the mutant Skf gene that shows a much larger amount of sporulation.

Figure 1b shows the mass of the eluted molecules to be consistent with those in DNA-dependent ATPase activity. Figure 1c shows enzymatic activity was only stimulated in the presence of duplex DNA. Figure 1d shows after cloning of D10 protein, D10-R389N (motif VI) was expressed and purified. When compared to wild type, a point mutation of the motif stopped all DNA-dependent ATPase activity.

Figure 1: Analyzed the growth curve based on cell density at 600nm (Figure 1C), and colony-forming unit calculations (Figure 1D) to identify the effect of gp70.1 (from the Bacteriophage PaP3) on P. aeruginosa PA3. The black circles on the graphs indicate the PA3 cells with the gp 70.1 gene; they had decreased cell density (1C) and decreased growth rate (1D); therefore, it was determined that Gp70.1 had a severe inhibitory effect on the concentration of bacterial culture.