 | Figure 2
Nature Biotechnology
20, 1140 - 1145 (2002)
Published online: 7 October 2002; | doi:10.1038/nbt747
Engineering tolerance and hyperaccumulation of arsenic in plants by combining arsenate reductase and  -glutamylcysteine synthetase expressionOm Parkash Dhankher, Yujing Li, Barry P. Rosen, Jin Shi, David Salt, Julie F. Senecoff, Nupur A. Sashti
& Richard B. Meagher | | | | Figure 2. Immunodetection of ArsC and  -ECS proteins in transgenic plants.
(A) Strong ArsC (16 kDa) protein expression was observed on western blots from leaves, but not roots of three independent lines transformed with the SRS1p/ArsC construct. Protein extracts from E. coli expressing ArsC from pNA1 plasmid and wild-type (WT) plant extracts serve as positive and negative controls, respectively. (B) Strong expression of ArsC protein was observed in both leaves and roots of three independent transgenic lines transformed with the 35Sp/ArsC construct. (C) Both  -ECS (top panel) and ArsC (bottom panel) are assayed on western blots of protein extracts from transgenic ArsC9 parental plant line expressing ArsC alone, a ACT2p/ECS1 line expressing g-ECS (57 kDa) alone, and ten lines generated by transforming the ArsC9 parental line with ACT2p/ -ECS (lines ArsC9 + ECS1−10). The western membrane was cut into two strips and reacted separately with ArsC- and -ECS-specific antisera. For (A−C), equal amounts (10  g) of total protein were resolved on a 12% (wt/vol) polyacrylamide gel by SDS−PAGE and blotted to membrane. Western blots of plant extracts were developed as described in Bizily et al.34 after reacting with polyclonal antisera to ArsC35 and monoclonal antibodies to -ECS31. Equal loading of samples was confirmed by Coomassie staining of parallel samples on a separate gel.
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