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

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.

authors

Koes R,Souer E,van Houwelingen A,Mur L,Spelt C,Quattrocchio F,Wing J,Oppedijk B,Ahmed S,Maes T

doi

10.1073/pnas.92.18.8149

subject

Has Abstract,Author List Incomplete

pub_date

1995-08-29 00:00:00

pages

8149-53

issue

18

eissn

0027-8424

issn

1091-6490

journal_volume

92

pub_type

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