Team:Warsaw/Calendar-Main/14 August 2008

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Cloning of alpha to pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_omega

Michał K.

Isolation of plasmids from cultures inocluated on previous day.
Digest of isolated plasmids with SacI (SacI buffer), vectors were also dephosphorylated with CIAP.
Gel electrophoresis and gel-out of proper bands 4400 bp (for pACYC177+OmpA_A_omega) and 4200 bp (pACYC177+OmpA_Z_omega).

Purification of proteins: Z-alpha and Z-omega

Piotr

Electrophoresis in polyacrylamide gel (12 %) of sonicate and preliminarily purified protein fusions: Z-alpha and Z-omega (Fig. 1. and Fig. 2.)

Fig. 1. Fractions from purifying Z-alpha and Z-omega protein fusions. 1. Purified Z-omega 2. Supernatant Z-alpha after sonication 3. Elution of Z-alpha using ddH2o 4. Purified Z-alpha 5. Marker

Fig. 2. Sonicate and pellet from Z-omega induction 1. Marker 2. Sonicate Z-omega after induction 3. Pellet Z-omega after induction