Figure 4.

Analysis of Columbia plants expressing the 35S:CsTFL transgene. A, Diagram of the CsTFL transgene including 35S CaMV promoter (black arrow), CsTFL cDNA (−65 to +587; gray arrow), and octopine synthase 3′-untranslated region (ocs 3′-UTR; black line). B, Detection of the 35S:CsTFL transgene in transformed Columbia plants using a 35S forward primer, a CsTFL gene-specific reverse primer, and genomic DNA in PCR reactions. Negative control for transgene detection was wild-type Columbia genomic DNA (WT Col). The positive control (+) was the pPSCsTFL-1 transgene construct. C, RNA blot analysis of 35S:CsTFL T₁ plants in wild-type Columbia background. Wild-type Columbia and representatives of three phenotypic classes of 35S:CsTFL T₁ plants were analyzed. For RNA blot analyses, RNA was isolated from a mixture of inflorescence and rosette leaves. Total RNA (1 μg) was loaded per lane, blotted, and hybridized with a ³²P-labeled CsTFL cDNA probe. The positive control (+) was 0.1 ng of in vitro-transcribed sense CsTFL RNA. The negative control was wild-type Columbia RNA. As a control for equal loading, a picture of the 25S ribosomal subunit from a gel stained with ethidium bromide is shown under the blots.