Analysis of the chromosomal distribution of transposon-carrying T-DNAs in tomato using the inverse polymerase chain reaction
1994
Thomas, C.M. (John Innes Centre, Norwich (United Kingdom). Sainsbury Lab.) | Jones, D.A. | English, J.J. | Carroll, B.J. | Bennetzen, J.L. | Harrison, K. | Burbidge, A. | Bishop, G.J. | Jones, J.D.G.
A system was developed afor isolating tomato genes by transposon mutagenesis. In maize and tobacco, the transposon Activator (Ac) transposes preferentially to genetically linked sites. To identify transposons linked to various target genes, the RFLP map locations of Ac- and Dissociation (Ds)-carrying T-DNAs were determined in a number of transformants. T-DNA flanking sequences were isolated using the inverse polymerase chain reaction (IPCR) and located on the RFLP map of tomato. The authenticity of IPCR reaction products was tested by several criteria including nested primer amplification, DNA sequence analysis and PCR amplification of the corresponding insertion target sequences. The RFLP map locations of 37 transposon-carrying T-DNAs is reported and also the map locations of nine transposed Ds elements. T-DNAs were identified on all chromosomes except chromosome 6. The data revealed no apparent chromosomal preference for T-DNA integration events. Lines carrying transposons at known map locations have been established which should prove a useful resource for isolating tomato genes by transposon mutagenesis
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