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Effect of deletions in the cauliflower mosaic virus polyadenylation sequence on the choice of the polyadenylation sites in tobacco protoplasts
Authors:François Guerineau  Louise Brooks and Philip Mullineaux
Institution:(1) AFRC Institute of Plant Science Research, John Innes Institute, NR4 7UH Colney Lane, Norwich, UK;(2) Present address: Scottish Crop Research Institute, DD2 SDA Invergowrie, Dundee, Scotland, UK
Abstract:Summary Deletions were made in the cauliflower mosaic virus polyadenylation sequence which was cloned downstream of the beta-glucuronidase gene (gus). The populations of mRNAs generated in tobacco mesophyll protoplasts by transient expression with the various constructs were analysed using a polymerase chain reaction procedure. When no deletion was present in the sequence, the mRNA appeared to be polyadenylated at two major polyadenylation sites. A deletion upstream from the AATAAA sequence made the population of polyadenylated mRNAs very heterogenous at their 3prime ends. A deletion downstream of the AATAAA sequence had no effect on the choice of the site. Alternative polyadenylation sites were used when the native polyadenylation site was deleted. These results are discussed in relation to data obtained with other polyadenylation sequences from both plants and animals.
Keywords:Polyadenylation  Gene expression  Cauliflower mosaic virus  Protoplasts  Polymerase chain reaction
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