Fig. 4:

Conventional Western blot (Panel I) of hydatid cyst fluid antigen B (arrowed) and plaque-dot blots of recombinant phages (Panel II) using individual sera from 26 mice (numbered at the top of Panel I) infected with oncospheres of E. granulosus. Recombinant phage vectors were transfected into XL1-Blue bacterial cells (Stratagene), then plated on top of LB plates with top agar. After growing the transfected cells for 5 h at 42°C, a nitrocellulose membrane was placed on top of the agar. Then the plate was incubated at 37°C for a further 4 h. The membrane was washed three times with PBST and cut into strips. After blocking in 5% (w/v) skim milk in PBS, the strips were reacted with the individual mouse sera and then sheep anti-mouse IgG conjugate (full details are described same as Western blotting in Protocol 2). The strips were developed with 4-chloro-1-naphthol substrate. Recombinant phage are EgTPx (a), EgG5 (b), EpC1 (c), and a mixture of EgTPx, EgG5 and EpC1 (d). The numbers of hydatid cysts resulting from oncospheral infection in each of the mice are shown between Panels I and Panels II. Protein standard markers (M) (in kDa) are shown on the left side of Panel I.