Figure 2

Strategy for subcloning the two BAC inserts and linking the BACs together BAC1 is recombined with pBACLinkSpAB linearized with NotI to give BAC1LinkSp. BAC2 is recombined with pBACLinkGmBC linearized with NotI to give BAC2LinkGm. BAC2LinkGm is linearized with I-PpoI and transfected into EL350 bacteria carrying BAC1LinkSp. Correctly recombined clones should be gentamycin resistant, chloramphenicol resistant and spectinomycin sensitive. The primers used to check each homologous recombination event are indicated as arrows labeled 1 to 5.