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Title of Journal: Plant Mol Biol Rep

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Abbravation: Plant Molecular Biology Reporter

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Springer-Verlag

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DOI

10.1016/0022-460x(90)90644-f

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1572-9818

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Isolation of cDNAs encoding for two distinct isofo

Authors: Pierre Vauclare Yasushi Kawagoe Norimoto Murai
Publish Date: 2012/09/03
Volume: 21, Issue: 4, Pages: 375-389
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Abstract

The TATAbox binding protein TBP is one of the 4 DNAbinding proteins that has been shown to associate with the proximal promoter region −295 of the gene for bean seed storage protein phaseolin The −295 promoter is essential for spatial and temporal control of the phaseolin gene expression We designed a pair of degenerated primers based on the highly conserved sequence of the carboxylterminal domain of yeast TBP and used PCR to amplify the corresponding sequence from the bean cDNA By using the amplified fragment as a probe we screened a cDNA library derived from poly A+ RNA from developing bean seeds and isolated 2 nearly fulllength cDNA clones 813 and 826 bp long The cDNAs encode 2 distinct isoforms of bean TBP PV1 and PV2 each with an open reading frame of 200 amino acid residues The 2 cDNA sequences share an 858 overall nucleotide sequence identity with the coding region showing a higher degree of identity 944 than the 5′ and 3′untranslated regions 69 The deduced amino acid sequence of the bean TBP isoforms differ in only 3 amino acid residues at positions 5 9 and 16 all located in the aminoterminal region The carboxylterminal domain of 180 amino acid residues shows a high degree 82 of evolutionary sequence conservation with the TBP sequences from other eukaryotic species This domain possesses the 3 highly conserved structural motifs namely the 2 direct repeat sequences a central basic region rich in basic amino acid residues and a region similar to the sigma factor of prokaryote On the basis of this and other findings we suggest that higher plants in general may have at least 2 copies of TBP gene presumably resulting from the global duplication of the genome

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