Abstract
Several clones corresponding to the gene encoding NADH-dependent hydroxypyruvate reductase have been isolated from a cucumber genomic library. Restriction mapping indicates the presence of two HPR genes, hpr-A and hpr-B, in the cucumber genome. Examination of the DNAs of individual plants suggests that hpr-A and hpr-B are most likely alleles at a single locus. The sequence of a 6.7 kb genomic fragment that includes the entire transcribed region, 2.2 kb of 5′ flanking sequence, and about 0.8 kb of 3′ flanking sequence reveals the presence of 12 introns in hpr-A. These introns are AT-rich relative to the exons. The donor sequence at the 5′ end of the sixth intron contains an unusual dinucleotide, GC, rather than the nearly invariant GT. Primer extension analysis maps the transcription initiation site to 61 nucleotides upstream of the translation initiation codon. An AT-rich stretch is centered at position −31 with respect to the transcription initiation site, and a potential CCAAT box is centered at position −138. Several elements that are homologous to regulatory elements of other plant genes have been identified in the flanking regions of hpr-A.
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References
Breathnach, R, Chambon, P: Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem 50: 349–383 (1981).
Brown, J: A catalogue of splice junction and putative branch point sequences from plant introns. Nucl Acids Res 14: 9549–9559 (1986).
Calzone, FJ, Britten, RJ, Davidson, EH: Mapping of gene transcripts by nuclease protection assays and cDNA primer extension. Meth Enzymol 152: 611–632 (1987).
Chomezynski, P, Sacchi, N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloro-form extraction. Analyt Biochem 162: 156–159 (1987).
Dellaporta, SL, Wood, J, Hicks, JB: A plant DNA miniprep: version II. Plant Mol Biol Rep 1: 19–21 (1983).
Feinberg, AP, Vogelstein, B: A technique for radiolabeling restriction endonuclease fragments to high specific activity. Analyt Biochem 132: 6–13 (1983).
Gilmartin, PM, Sarokin, L, Memelink, J, Chua, N-H: Molecular light switches for plant genes. Plant Cell 2: 369–378 (1990).
Greenler, JM, Becker, WM: Organ specificity and light regulation of NADH-dependent hydroxypyruvate reductase transcript abundance. Plant Physiol 94: 1484–1487 (1990).
Greenler, JM, Sloan, JS, Schwartz, BW, Becker, WM: Isolation, characterization and sequence analysis of a fulllength cDNA clone encoding NADH-dependent hydroxypyruvate reductase from cucumber. Plant Mol Biol 13: 139–150 (1989).
Henikoff, S: Unidirectional digestion with exonuclease III in DNA sequence analysis. Meth Enzymol 155: 156–165 (1987).
Hondred, D, Wadle, D-M, Titus, DE, Becker, WM: Light-stimulated accumulation of the peroxisomal enzymes hydroxypyruvate reductase and serine:glyoxylate aminotransferase and their translatable mRNAs in cotyledons of cucumber seedlings. Plant Mol Biol 9: 259–275 (1987).
Husic, DW, Husic, DH, Tolbert, NE: The oxidative photosynthetic carbon cycle or C2 cycle. Crit Rev Plant Sci 5: 45–100 (1987).
Jacob, M, Gallinaro, H: The 5′ splice site: phylogenetic evolution and variable geometry of association with U1RNA. Nucl Acids Res 17: 2159–2180 (1989).
Kaiser, K, Murray, NE: The use of phage Lambda replacement vectors in the construction of representative genomic DNA libraries. In Glover, DM (ed) DNA Cloning, vol 1, pp. 1–47. IRL Press, Washington, DC (1985).
Kuhlemeier, C, Green, PJ, Chua, N-H: Regulation of gene expression in higher plants. Annu Rev Plant Physiol 38: 221–257 (1987).
Maniatis, T, Fritsch, EF, Sambrook, J: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).
Sanger, F, Coulson, AR, Barrell, BG, Smith, AJH, Roe, BA: Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143: 161–178 (1980).
Schindler, U, Cashmore, AR: Photoregulated gene expression may involve ubiquitous DNA binding proteins. EMBO J 9: 3415–3427 (1990).
Tobin, EM, Silverthorne, J: Light regulation of gene expression in higher plants. Annu Rev Plant Physiol 36: 569–593 (1985).
Watson, JC, Thompson, WF: Purification and restriction endonuclease analysis of plant nuclear DNA. Meth Enzymol 118: 57–75 (1986).
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Schwartz, B.W., Sloan, J.S. & Becker, W.M. Characterization of genes encoding hydroxypyruvate reductase in cucumber. Plant Mol Biol 17, 941–947 (1991). https://doi.org/10.1007/BF00037078
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DOI: https://doi.org/10.1007/BF00037078