Genomic organization and transcriptional analysis of STDEFICIENS in Solanum tuberosum L.
The genomic organization of STDEFICIENS (STDEF), the potato orthologous gene to DEFICIENS (DEF) from Antirrhinum majus and APETALA3 (AP3) from Arabidopsis thaliana, has been investigated. Southern-blot analysis on genomic DNA from dihaploid potato lines, using 5'-gene specific probes, revealed polymorphisms that were consistent with the existence in potato of at least two copies of STDEF per haploid genome. This was confirmed by the detection of at least six different STDEF transcripts in the common tetraploid potato S. tuberosum. Genes for two of the STDEF loci, here designated as STDEF-1 and STDEF-2, have been identified as corresponding to the previously described pD13 and pD12 genomic clones, respectively (García-Maroto et al., 1993). In addition we have characterised the transcriptional STDEF unit. The main transcription start has been identified around 90 nt upstream of the putative initiation ATG codon, at a CAAATC motif, conserved in AP3. An additional transcription initiation site was detected by 5'-RACE analysis about 300 nt upstream of the main start, which has been confirmed by reverse transcriptase - polymerase chain reaction (RT-PCR) amplification from the longer transcripts. A comparison of the promoter regions for pD12, pD13 and AP3 indicates a similar overall structure, but reveals the existence of a great divergence between pD12 and pD13 in a promoter region that should contain important cis-regulatory elements. This raises the possibility of a differential regulation for the two STDEF genes.
Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos, Universidad Politécnica de Madrid, Avda, Complutense s/n, E-28040 Madrid, Spain. fgmaroto@ual.es
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García-Maroto
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MJ
Carmona
D001483:Base Sequence; D015139:Blotting, Southern; D026281:DEFICIENS Protein; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004171:Diploidy; D017343:Genes, Plant / Q000235:genetics*; D005838:Genotype; D018398:Homeodomain Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011123:Polyploidy; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D011198:Solanum tuberosum / Q000235:genetics*; D014158:Transcription, Genetic
D026281:DEFICIENS Protein; D018076:DNA, Complementary; D018744:DNA, Plant; D018398:Homeodomain Proteins; D012333:RNA, Messenger