Team:Warsaw/Calendar-Main/13 August 2008

Checking for presence of protein A on the cell membrane

Piotr

 Pouring bacteria by drops onto nitrocellulose. Drying. Blocking. Anti-A antibody binding. Washing. Anti-rabbit antibody binding. Developing with BCIP and NBT (see Fig. 1.).</li> </ol> <img src="http://2008.igem.org/wiki/images/e/ec/Blot_OmpA_WAW.jpg" width=150/></a> Fig. 1.Protein A detection on the surface of bacteria. 1 - E. coli expressing OmpA_Omega_A_Alpha, 2 - E. coli expressing OmpA_Z_Omega (antibodies binds to protein Z probably due to similarity of protein A and Z), 3 - E. coli expressing OmpA_A_Alpha, 4 - E. coli carrying OmpA_Omega_A_Alpha without inducer, 5 - E. coli expressing OmpA_A_Omega, 6 - E. coli Top10 without plasmid.

Cloning of alpha to pACYC177+OmpA_A_omega</a> and pACYC177+OmpA_Z_omega</a> Michał K.  Inoculation of pACYC177+OmpA_A_omega</a> and pACYC177+OmpA_Z_omega</a> plasmids to LB with kanamycin.