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Nucleic Acids Research, 1989, Vol. 17, No. 23 9993-10013
© 1989

MOLECULAR BIOLOGY

The complete sequence of soybean chlorotic mottle virus DNA and the identification of a novel promoter

A. Hasegawa1, J. Verver+, A. Shimada1, M. Saito1, R. Goldbach§, A. Van Kammen+, K. Miki1, M. Kameya-Iwaki2 and T. Hibi*

Laboratory of Applied Microbiology, National Institute of Agrobiological Resources Tsukuba Science City, Kannondai, Ibaraki 305 1Fundamental Research Laboratory TONEN Co., 1-3-1 Nishitsurugaoka, Ohi-machi, Iruma-gun, Saitama-ken 354 2Department of Plant Protection, National Agricultural Research Center Tsukuba Science City, Kannondai, Ibaraki 305, Japan

*To whom correspondence should be addressed

Received July 28, 1989. Revised October 24, 1989. Accepted October 24, 1989.

The complete nucleotide sequence of an infectious clone of soybean chlorotic mottle virus (SoyCMV) DNA was determined and compared with those of three other caulimoviruses, cauliflower mosaic virus (caMV), carnation etched ring virus and figwort mosaic virus. The double-stranded DNA genome of SoyCMV (8,175 bp) contained nine open reading frames (ORFs) and one large intergenic region. The primer binding sites, gene organization and size of OREs were similar to those of the other caulimoviruses, except for ORF I, which was split into ORF Ia and lb. The amino acid sequences deduced from each ORF showed only short, highly homologous regions in several of the corresponding ORFs of the three other caulimoviruses. A promoter fragment of 378 bp in SoyCMV ORF III showed a strong expression activity, comparable to that of the CaMV 35S promoter, in tobacco mesophyll protoplasts as determined by a ß-glucuronidase assay using electrotransfection. The fragment contained CAAT and TATA boxes but no transcriptional enhancer signal as reported for the CaMV 35S promoter. Instead, it had sequences homologous to a part of the translational enhancer signal reported for the 5'-leader sequence of tobacco mosaic virus RNA.

+Department of Molecular Biology Agricultural University, De Dreyen 11, 6703 BC Wageningen, The Netherlands

§Department of Virology, Biology Agricultural University, De Dreyen 11, 6703 BC Wageningen, The Netherlands


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