FIG. 2.

(A) Physical and genetic map of the region containing raiI and raiR from R. etli CNPAF512. Coding regions are shaded. Arrows indicate directions of transcription. The open arrow represents the beginning of an open reading frame. Light shading illustrates a part of ORF3 that has not been sequenced. The boxed SalI sites have been used for construction of pFAJ1323. The small arrows indicate the primers with NotI sites used for amplification of a 4-kb fragment. Restriction sites: B, BamHI; H, HindIII; P, PstI; S, SphI; Sl, SalI; Sm, SmaI. (B) Construction of the raiI (FAJ1328) and raiR (FAJ1329) mutant. The PCR product was cloned into pCR2.1 TA, excised with NotI, and inserted into pJQ200uc1. A cassette with a promoterless gusA and kanamycin resistance from pWM6 was introduced in the XhoI (raiI mutant) or NdeI (raiR mutant) site by blunt-end ligation.