Fig. 1

Deletion of glycosyltransferase in C. bescii. a Chromosome map of the glycosyltransferase (Cbes1864) and surrounding genes. b Depiction of the deletion cassette consisting of a fused 5′ and 3′ flanking region in a non-replicating plasmid, pJRW012, with a copy of the pyrF gene from Clostridium thermocellum (Clo1313_1266) for selection of uracil prototrophic transformants of a ΔpyrF background strain. Counterselection with 5-FOA selected strains that had undergone a second recombination event resulting in strain JWCB143 (ΔpyrF ΔcbeI Δcbes1864). c Agarose gel showing PCR products amplified with primers SK163 and SK164 on gDNA templates from wild-type C. bescii (lane 3), parent strain JWCB018 (lane 4), Δcbes1864 strain JWCB143 (lane 5), and no template (lane 2). Lane 1: DNA molecular weight standards, Lanes 6–8: JWCB143 sister isolates. Expected band for wild-type locus is 3.7 kb and for cbes1864 deletion locus is 2.2 kb