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9 cfu/ml. The full-length and delted AvrPto constructs were cloned into the pBIN19 derivative pMD1. Positive clones were introduced into Agrobacterium via electroporation.
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Mutations were introduced to the Pto gene sequence by recombinant polymerase chain reaction with the use of cloned Pto and Fen templates (D. T. L., J.P.R. and J.H.C., unpublished data). Recombinant PCR products were cloned into the yeast two-hybrid vector pAS2-1 (Clontech) and sequenced before testing. Wild-type and mutant avrPto genes were cloned into a pACT2 (Clontech).
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We thank D. O. Lavelle, J. Gardner, and J. Luke for DNA sequencing and G. Oldroyd, M. B. Mudgett, S. Barker, V. Szabo, and L. Doyle for comments on the manuscript. Supported by NSF Cooperative Agreement BIR-8920216 to CEPRAP and by CEPRAP corporate associates Calgene, Inc., Ciba Geigy Biotechnology Corporation, Sandoz Seeds, and Zeneca Seeds.
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