Cloning of truncated fragment of protein A (ΔA)

Piotr

Inoculation of some pACYC177+OmpA_alpha + ΔA and pACYC177+OmpA_omega + ΔA colonies of tranformants.

Checking the expression of OmpA_omega_ΔA_alpha and OmpA_A_alpha

Piotr

Inoculation of OmpA_omega_ΔA_alpha and OmpA_A_alpha with inductor (0,5 mmol/mL IPTG) and negative control without IPTG.

Checking if OmpA_omega_ΔA_alpha gives ampicillin resistance

Emilia

Inoculation of OmpA_omega_ΔA_alpha into various IPTG concentrations: 0, 0.1, 0.25, 0.5, 0.75, 1 mmol/mL.

Preparing pACYC177+OmpA_omega_ΔA construct

Michał K.

1. Digest of pACYC177+OmpA_omega_ΔA_alpha (from 25 July) with BamHI and NotI (BamHI buffer). DNA ends blunting with Klenow fragment (3 hr).
2. Gel elctrophoresis (Fig. 1) and gel-out of proper band - 4300 bp.
3. Overnight ligation of isolated DNA fragment.