Tests for ampicillin resistance given by protein added to mediumPiotr
Results of growth from previous day
OmpA variant ("hunter") | "Prey" variant | Growth with prey | Growth without prey |
OmpA-alpha | His+Z+omega | + | - |
OmpA-alpha | His+Z+alfa | - | - |
OmpA-A-alpha | His+Z+omega | + | - |
Cloning of protein A DNA to pET15b+OmpA-alpha plasmidMichał K.
- Digest of pGeneart+A and pET15b+OmpA-alpha with NdeI and NotI (Orange buffer), pET15b was also dephosphorylated.
- Gel electrophoresis of digested plasmids (Fig. 1.) and gel-out of proper bands (400 bp for A lane and 6200 bp for pET15b lane).
- Ligation of pET15b-Alpha and A.
- Transformation of E. coli TOP10.
- Transformants plating on LB + ampicillin.
Fig. 1. NdeI/NotI digest of plasmid pGeneart+A
1. Marker
2. Digested plasmid pGeneart+A
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