Figures:
Figure 1. Development and characterization of CTB-TM-1 expression cassette in transgenic wheat seeds. A. The schematic diagram of expression cassette of pPZP211: CTB-TM-1. Genetic components included are: PGluB-4, promoter of glutelin gene GluB-4; CTB, Cholera non-Toxin B subunit; Furin, Furin cleavage site; TM-1, TM-1 protein ofMycoplasma gallisepticum (MG); KDEL, Endoplasmic reticulum retention signal; Tnos, termination of the nopaline synthase gene fromAgrobacterium tumefaciens; CaMV 35S, promoter of 35S RNA fromCauliflower Mosaic Virus ; KanR, Kanamycin resistant gene; CaMV Poly(A), Cauliflower Mosaic Viruspolyadenylation signal; LB and RB, left and right border of T-DNA repeat respectively. Arrows indicate PCR and sequencing primers. B.characterization of genomic integration of pPZP211: CTB-TM-1 cassette in different transgenic lines by PCR using primers (CTB-F and TM-1-R).C. Validation of three independent lines for recombinant CTB-TM-1 expression by Western blot with anti-CTB antibody. D.quantification of recombinant CTB-TM-1 expression in C. E. GM1 ELISA detecting the folding of CTB-TM-1 and its binding with GM1 receptor. *, P <0.05, **, P <0.01, ***,P <0.001.