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Versatile transformation vectors to assay the promoter activity of DNA elements in plants
G Pasquali1, P B Ouwerkerk, J Memelink
1Institute of Molecular Plant Sciences, Leiden University, Clusius Laboratory, The Netherlands.
Abstract:
A convenient vector system was developed to evaluate transcriptional promoter activities in plants. Two primary vectors, optionally containing the cauliflower mosaic virus (CaMV) 35S -47 or -90 minimal promoters, offer multiple sites for cloning the sequence of interest upstream from the beta-glucuronidase gene (gusA). The promoter-gusA cassette can be transferred to a binary vector containing the selectable neomycin phosphotransferase II-encoding gene (nptII) next to the left border. In addition, the transferred DNA (T-DNA) contains the chloramphenicol acetyltransferase gene (cat) driven by the CaMV 35S promoter. Activity of cat can serve as a reference for gusA expression to correct for effect of chromosomal position or T-DNA copy number.