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Nucleic Acids Research, 1983, Vol. 11, No. 19 6821-6835
© 1983

MOLECULAR BIOLOGY

Construction of infectious potato spindle tuber viroid cDNA clones

Dean E. Cress1, Michael C. Kiefer3 and Robert A. Owens2

1Tissue Culture and Molecular Genetics Laboratory USA 2Plant Virology Laboratory, US Department of Agriculture Beltsville, MD 20705, USA 3Department of Botany, University of Maryland College Park, MD 20742, USA

Received March 21, 1983. Revised September 8, 1983. Accepted September 8, 1983.

Contiguous restriction fragments from two cloned partial-length potato spindle tuber viroid (PSTV) cDNAs were used to construct recombinant DNAs containing full-length monomeric and dimeric PSTV cDNA. When five different PSTV cDNA plasmids and RNA isolated from E. coli cells harboring these plasmids were tested for infectivity on tomato, plasmid DNAs containing PSTV cDNA dimers were infectious. RNA transcripts containing the sequence of PSTV from these plasmids were also infectious. The sequences of the viroid progeny and the cloned DNA were identical. In vitro mutagenesis of infectious PSTV cDNAs will allow systematic investigation of the role of specific sequences in viroid replication and pathogenesis.


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