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Using Mu killer

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For data on the efficiency of silencing of a multiple MuDR transposon tagging line by a Muk heterozygote, please see the Slotkin et al., Genetics publication. PDF 350K

For information on how Mu killer can effect plant phenotype, download this PDF (2.1M) figure.

For information on how to reduce the copy number of Mutator transposons, download this PDF (252K) figure.

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How efficient is Mu killer at silencing a typical Mutator tagging line?

To test the efficiency of MuDR silencing by Mu killer, we crossed a single individual used for Mutator transposon tagging with multiple active MuDR elements to both a Mutator permissive tester, and homozygous Mu killer. We grew equal amounts of all seed phenotypic classes from the progeny of these crosses. DNA from the progeny was subjected to Southern analysis with HinfI digestion and probed with the internal region of Mu1.

Active Mutator individuals have only the smaller 1.4kb band, while individuals with inactive MuDR transposons have hypermethylated Mu1 TIRs and bands that ladder larger than 1.4kb. A subset of this data is shown below. Nearly 100% of individuals crossed to homozygous Mu killer result in a silenced Mutator family.

NOTE: DNA used for the analysis shown below is from young seedling leaves. Individuals with silencing MuDR elements that have both the active 1.4kb band and larger laddering bands are in the process of silencing. The inactivation of the Mutator family is complete by floral development.