Figure 2. trans-Complementation of the rpoS mutant and modification of the mutant to constitutively express OspC and to simultaneously express OspC, DbpA and DbpB.

(A&B) Construction of pBBE22-rpoS and pME22-C′B′A′. All restriction enzyme sites and primer binding sites used for plasmid construction are labeled. (C) Confirmation of OspC, DbpA and DbpB production by immunoblotting. The parental clone 13A and transformants ΔrpoS/E22, ΔrpoS/rpoS, ΔrpoS/ospC′, ΔrpoS/C′B′A′ and 13A/C′B′A′ were grown to late-log phase in BSK-H complete medium, harvested by centrifugation and subjected to immunoblot analyses probed with a mixture of FlaB mAb and OspC mAb (top), or mouse anti-DbpA (middle) or -DbpB sera (bottom).