Chromosomal integration of the E1 gene into the C. bescii genome. a A diagram of native E1 protein: SP, signal peptide; a family 5 endoglucanase; and CBM2, a family 2 carbohydrate-binding module/domain. The black bar beneath the diagram represents the portion of the pDCW175 construct derived from Acel_0614. b A depiction of the chromosomal location and integration event of the E1 expression cassette/secretion cassette. c Agarose gel showing PCR products amplified using primers DC477 and DC478 annealing to regions outside the site of integration in the newly constructed strain JWCB052 ΔpyrFA ldh::ISCbe4 Δcbe1::PS-layer
acel0614 E1 containing the E1 expression cassette, 4.079 kb (lane 2) and the parent strain, JWCB018 ΔpyrFA
ldh::ISCbe4 Δcbe1, 2.440 kb (lane 3); DNA MW standards (lane 1); no template PCR control (lane 4).