Meiotically Stable Natural Epialleles of Sadhu, a Novel Arabidopsis Retroposon (original) (raw)
Figure 4
RT-PCR/CAPS (Cleaved Amplified Polymorphic Sequence) Detection of Allele-Specific Expression at At2g10410
The Col allele is cleaved with _Bst_B1, generating a 480-bp fragment plus an undetected 70-bp fragment, while the Ler allele is uncleaved (550 bp). At2g10410 is not expressed in Ler wild-type, but is expressed in wild-type Col and Ler ddm1 mutant, as shown in Figure 1. A mixture of cDNA templates from Col and Ler ddm1 samples was used to illustrate the detection of bi-allelic expression; note an additional higher molecular weight heteroduplex band (see Materials and Methods) in samples showing bi-allelic expression. A total of five Col × Ler heterozygous At2g10410 BC1 and ten Col × Ler ddm1–2 heterozygous BC1 individuals were examined; all looked identical to the representative individuals shown. In addition, we examined two DDM1+/+ F2 individuals homozygous for the Ler allele at At2g10410 that resulted from self-pollination of a Ler ddm1 × Col F1 individual. RT-PCR amplification of cyclophilin transcripts is shown as a control.