Figure 3
Presence and size of multiple promoter:bovine lysozyme (BvLzm)-terminator cassettes in the same BvLzm transgenic line as determined by PCR analysis. Representative lines with single or multiple promoter:BvLzm-terminator cassettes are shown. (1) pU:BvLzm-35ST line; (2) pUD:BvLzm-35ST line; (3) pUDE:BvLzm-3′UTR-35ST line; (4) pUPE:BvLzm-3′UTR-35ST line; (5) pUPBE:BvLzm-3′UTR-35ST line; (6) pUPBE:BvLzm-35STNOST line; (7) vector-transformed line; (8) non-transformed (NT; tissue culture-derived) plant; and (9) no DNA template (negative control for PCR). (a) Detection of pUbi, BvLzm, 3′UTR, 35ST and NOST using the primer sets pUbi-F/35ST-R (2.62 kilobase pairs [kb] or 2.85 kb fragment) and pUbi-F/NOST-R (2.87 kb fragment). (b) Detection of pSHDIR16, BvLzm, 3′UTR, 35ST and NOST using the primer sets pSHDIR16-F/35ST-R (3.32 kb fragment) and pSHDIR16-F/NOST-R (3.56 kb fragment). (c) Detection of pSHPRP, BvLzm, 3′UTR, 35ST and NOST using the primer sets pSHPRP-F/35ST-R (3.65 kb fragment) and pSHPRP-F/NOST-R (3.90 kb fragment). (d) Detection of pSHEF1α, BvLzm, 3′UTR, 35ST and NOST using the primer sets pSHEF1α-F/35ST-R (2.57 kb fragment) and pSHEF1α-F/NOST-R (2.82 kb fragment). (e) Detection of pSCBV21, BvLzm, 3′UTR, 35ST and NOST using the primer sets pSCBV21-F/35ST-R (2.21 kb fragment) and pSCBV21-F/NOST-R (2.46 kb fragment). BvLzm, maize codon-optimized bovine lysozyme gene; U, Ubi promoter; D, SHDIR16 promoter; P, SHPRP promoter; E, SHEF1α promoter; B, SCBV21 promoter; 3′UTR, 3′ untranslated region of Sorghum mosaic virus. 35ST, Cauliflower mosaic virus 35S terminator; NOST, Agrobacterium tumefaciens nopaline synthase terminator. Full-length uncropped gels of the PCR products are displayed in Supplementary Figures S4, S5 and S6.
