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PMID: 3860847 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

T-DNA border sequences required for crown gall tumorigenesis.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 82 ·No. 15 ·1985-08-00 ·Pages 5112-6

Peralta EG, Ream LW

Abstract

Similar 23-base-pair (bp) direct repeats occur at the ends of two adjacent but noncontiguous T-DNAs, TL and TR (left and right T-DNA), in the tumor-inducing plasmid pTiA6NC. Thus, three border repeats lie right and one lies left of TL, which carries the genes needed for tumor maintenance. To determine whether T-DNA transfer and integration (subsequently called T-DNA transmission) require sequences in addition to the 23-bp border repeat, we constructed a deletion removing the three potential TL right borders (the TL right border and both TR borders). Since this deletion severely attenuated virulence, we reintroduced restriction fragments containing the TL right border repeat at a new location to the right of TL and tested their ability to restore virulence. Fragments that carried the border repeat flanked by at least 67 bp of wild-type Ti plasmid sequences on the left and 1035 bp on the right restored virulence completely. Smaller fragments restored virulence significantly but not fully, even though the border repeat remained intact. Therefore, T-region sequences flanking the border repeat in the fully active fragments stimulated T-DNA integration. Fragments that restored virulence fully when inserted in the wild-type orientation stimulated virulence only slightly in the opposite orientation. Thus, the right border sequence promotes T-DNA transfer and integration best in one direction.

MeSH Terms
Base Sequence Chromosome Deletion DNA, Bacterial/genetics Gene Expression Regulation Plant Tumors/microbiology Plants Plasmids Repetitive Sequences, Nucleic Acid Rhizobium/genetics
Chemicals
DNA, Bacterial
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Peralta E G
Ream L W
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35 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.82.15.5112
Published
1985-08-00
Pages
5112-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC390509
Subset
IM
Grants
NCRR NIH HHS · S07 RR 7031-F · United States
External Links
PubMed source DOI: 10.1073/pnas.82.15.5112 PMC Full Text View journal details
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