Cloning of protein Z DNA to OmpA constructs

Michał K.

1. Digest and dephosphorylation (with CIAP) of pACYC177+OmpA_alpha with SacI and NotI (BamHI buffer).
2. Gel eloctrophoresis and gel-out (4300 bp band).
3. Ligation of isolated DNA fragment and Z DNA fragment isolated on 14 July.
4. Transformation of E. coli TOP10 strain with ligation.
5. Transformants plating on LB + kanamycin.

Cloning of protein Z DNA to pET15b-OmpA-omega in place of OmpA

Paweł

1. Digest of pET15b-OmpA-omega and Z(in GeneArt vector) with NdeI and NotI.
2. Gel-out of Z (~200 bp band).
3. Overnight ligation of Z into digested pET15b-OmpA-omega.