Journal: Plant Physiology
Article Title: Dual Temporal Role of Plastid Sigma Factor 6 in Arabidopsis Development 1Dual Temporal Role of Plastid Sigma Factor 6 in Arabidopsis Development 1 [OA]
Figure Lengend Snippet: Characterization of the Arabidopsis sig6 - 2 mutant. A, Genomic Sig6 region (At2g36990) showing exon/intron structure and T-DNA insertion site in exon 5, 1,022 nucleotides downstream from the ATG start codon (top line). The resulting cDNA with the fused exons, but without the 5′ and 3′ untranslated regions, is depicted below. Also given are features of the derived protein. TP, Transit peptide; UR, unconserved region; 1–4, conserved regions for sigma activity. Genomic sequence and cDNA, but not T-DNA, are drawn to scale (scale bar on top). LB, Left border; RB, right border; sul , sulfonamide (sulfadiazine) resistance gene. B, RT-PCR detection of sigma factor transcripts in wild type (WT) and sig6 - 2 mutant. Total RNA was prepared from 6-d seedlings, reverse transcribed, and cDNA was amplified using the gene-specific primer pairs as described in “Materials and Methods.” C, Wild-type (WT) and sig6 - 2 mutant phenotype during development (4, 8, 10, 12, 21, and 28 d after sowing).
Article Snippet: Total RNA (2 μ g) from 6-d-old Arabidopsis seedlings was mixed with random primers (10 pM; Promega), incubated at 70°C for 10 min, and chilled on ice for 1 min. After addition of 6 μ L avian myeloblastosis virus-reverse transcriptase buffer (Promega), 1 μ L RNasin (40 units/ μ L; Promega), 3 μ L dNTPs (0.25 m m each), and 3 μ L avian myeloblastosis virus-reverse transcriptase (10 units/ μ L) to a final volume of 30 μ L, the reaction was incubated at 37°C for 90 min.
Techniques: Mutagenesis, Derivative Assay, Activity Assay, Sequencing, Reverse Transcription Polymerase Chain Reaction, Amplification