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  • Title: Targeted gene inactivation in petunia by PCR-based selection of transposon insertion mutants.
    Author: Koes R, Souer E, van Houwelingen A, Mur L, Spelt C, Quattrocchio F, Wing J, Oppedijk B, Ahmed S, Maes T.
    Journal: Proc Natl Acad Sci U S A; 1995 Aug 29; 92(18):8149-53. PubMed ID: 7667260.
    Abstract:
    Establishment of loss-of-function phenotypes is often a key step in determining the biological function of a gene. We describe a procedure to obtain mutant petunia plants in which a specific gene with known sequence is inactivated by the transposable element dTph1. Leaves are collected from batches of 1000 plants with highly active dTph1 elements, pooled according to a three-dimensional matrix, and screened by PCR using a transposon- and a gene-specific primer. In this way individual plants with a dTph1 insertion can be identified by analysis of about 30 PCRs. We found insertion alleles for various genes at a frequency of about 1 in 1000 plants. The plant population can be preserved by selfing all the plants, so that it can be screened for insertions in many genes over a prolonged period.
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