Team:Warsaw/Calendar-Main/14 August 2008

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</a> of proper bands 4400 bp (for pACYC177+OmpA_A_omega</a>) and 4200 bp (pACYC177+OmpA_Z_omega</a>). </li></ol>

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.</a> and Fig. 2.</a>)

<img src="http://2008.igem.org/wiki/images/3/35/August_14_th_2.jpg" width=250 /></a> 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

<img src="http://2008.igem.org/wiki/images/5/5d/August_14_th.jpg" width=220 /></a> Fig. 2. Sonicate and pellet from Z-omega induction 1. Marker 2. Sonicate Z-omega after induction 3. Pellet Z-omega after induction