Skip to main content
Figure 5 | BMC Plant Biology

Figure 5

From: Identification of precursor transcripts for 6 novel miRNAs expands the diversity on the genomic organisation and expression of miRNA genes in rice

Figure 5

Identification and validation of osa-miR827a and osa-miR1874 transcript precursors. A) Schematic for cloning of pre-osa-miR827a and pre-osa-miR1874 by cRT-PCR. The sequence was cloned by cRT-PCR from total RNA extracted from rice seedlings, as indicated in methods. Arrows indicated by R and F refers to primers for PCR amplification steps. The gel shows analysis of of cRT-PCR amplified products products. - and + refers to absence or presence of reverse transcription previous to PCR steps, as a control for DNA contamination. B) The cloned cRT-PCR products and folding into stem-loop structures. 5' and 3' end indicates the extremities of the cloned products. C) Transient expression of pre-osa-miR827a and pre-osa-miR1874 in Nicotiana benthamiana leaves. The predicted stem-loop encompassing miR827a or miR1874 plus 150 nucleotides of 5' and 3' sequences was expressed from pCUbi vector in Nicotiana benthamiana IR lines leaves, as schematised. P::Ubizm indicates maize Ubiquitin promoter and T:: Nos indicates Nopaline synthetase polyA site. The products were analysed by Northern blot using oligonucleotide probes complementary to osa-miR827a or osa-miR1874. Mock is a control of leaves transfected with an empty pcUbi vector.

Back to article page