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Nucleic Acids Research, 1985, Vol. 13, No. 2 459-471
© 1985

Articles

Unusual plasmid DNA organization in an octopine crown gall tumor

William W. Kwok1 and Eugene W. Nester+

Department of Biochemistry, University of Washington Seattle, WA 98195, USA +Department of Microbiology and Immunology, University of Washington Seattle, WA 98195, USA

Received September 21, 1984. Revised December 11, 1984. Accepted December 11, 1984.

A cloned tobacco crown gall tumor, 1595501, incited by A. tumefaciens strain 15955 was studied. Molecular analysis of the organization of the T-DNA by means of Southern transfer and hybridization techniques indicated that the 1595501 tumor has about 10 copies of TL DNA, five of which are complete TL DNA, whereas most octopine tumors have only one to two copies of complete TL DNA. Hybridization studies and genomic cloning indicated that some segments of the T-DNA have undergone deletions. One of the clones contained two copies of T-DNA which were inverted in orientation with respect to each other.

Two left ends of TL DNA from the 1595501 tumor line and the corresponding region of the octopine plasmid were sequenced. Comparison of the various cloned T-DNA sequences with Ti-plasmid sequence indicated that while there is an association with a 25 base pair direct repeat, there is no specific set of base pairs in the T-DNA at which divergence from Ti-plasmid sequences occurred.

1Present address: Fred Hutchinson Cancer Research Center, 1124 Columbia Street, Seattle, WA 98104, USA


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