Team:Warsaw/Calendar-Main/13 August 2008

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Checking for presence of protein A on the cell membrane

Piotr

  1. Pouring bacteria by drops onto nitrocellulose.
  2. Drying.
  3. Blocking.
  4. Anti-A antibody binding.
  5. Washing.
  6. Anti-rabbit antibody binding.
  7. Developing with BCIP and NBT (see Fig. 1.).

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 and pACYC177+OmpA_Z_omega

Michał K.

Inoculation of pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega plasmids to LB with kanamycin.

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