Supplemental Data Cloning, Expression, and Biochemical Characterization of Streptomyces rubellomurinus Genes Required for Biosynthesis of Antimalarial Compound FR900098

coli DH10B derivative that contains the trfA33 gene under the control of a rhamnose-inducible promoter, which, when induced by addition of 20 mM rhamnose, allows replication of pJK050 as a high-copy plasmid. WM4489 ((mcrA, ∆(mrr, hsdRMS, mcrBC), φ80(∆lacM15), ∆lacX74, endA1, recA1, deoR, ∆(ara, leu)7697, araD139, galU, galK, nupG, rpsL, λattB::pAE12(PrhaB::trfA33, ∆oriR6K-cat::frt5)) is similar to WM3118, except that the antibiotic resistance marker has been deleted. E. coli WM3608(Blodgett et al., 2005) was used as a donor strain for conjugal transfer of plasmids into S. lividans. Plasmids used for cloning in the study are shown in Figure S2. The biosassay strain E. coli WM6242 (lacIq, rrnB3, del(lacZ4787), hsdR514, attP22(EcoB), del(araBAD)567, del(rhaBAD)568, rph-1, del(phnC-P), del(phoA), HK-attB::pJK077(del aadA-oriR6K), lambda-attB::pJK074(delcat-oriR6K) expresses the phosphonate uptake system encoded by phnCDE under the control of a Ptac promoter. Induced expression of phnCDE by addition 0.5 mM IPTG increases sensitivity to many phosphonate antibiotics. WM6242 is a derivative of BW26678(Haldimann and Wanner, 2001). The plasmids containing phnCDE, pJK074 and pJK077, are derivatives of pAE12