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. 1995 Aug 29;92(18):8149–8153. doi: 10.1073/pnas.92.18.8149

Targeted gene inactivation in petunia by PCR-based selection of transposon insertion mutants.

R Koes 1, E Souer 1, A van Houwelingen 1, L Mur 1, C Spelt 1, F Quattrocchio 1, J Wing 1, B Oppedijk 1, S Ahmed 1, T Maes 1, et al.
PMCID: PMC41113  PMID: 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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Selected References

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