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The T-strand, a probable intermediate of Agrobacterium plant transformation, is bound by a nondenaturable linkage to a protein moiety at its 5′ end. The protein is shown to be the polypeptide VirD2, previously identified as a component of the T-DNA border endonuclease that initiates T-strand production. T-strands from an Agrobacterium strain expressing a virD2-lacZ fusion are bound to a protein of larger size than the wild-type protein and are immunoprecipitable by antibody to β-galactosidase.