Summary
The nucleotide sequence of gz19ab11, a clone that corresponds to the coding and flanking sequences of an Mr 19000 alpha zein, was determined. Comparison of the DNA sequences flanking this gene with those of other members of the gene subfamily showed that sequence conservation extends 820 nucleotides into the 5′ flanking region and 130 nucleotides into the 3′ flanking region. Southern blot analysis of maize DNA indicated that highly repetitive sequences are located within 950 bp 5′ and 300 bp 3′ to the protein coding region of these genes. The coding region of gz19ab11 is similar to but not identical with cDNA clones corresponding to Mr 19000 zeins, and analysis of zein transcripts indicated that this gene is expressed exclusively in endosperm tissue. RNAs which correspond to transcripts originating 60 nucleotides, and more than 800 nucleotides, upstream of the initiation codon were detected for this and a related gene. However, the concentration of the large RNA species was several orders of magnitude less than that of the shorter RNAs. The functional significance of these large RNA transcripts in zein gene expression is unclear.
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References
Benoist C, O'Hare K, Breathnach R, Chambon P (1980) The ovalbumin gene—sequence of putative control regions. Nucleic Acids Res 8:127–142
Benton WD, Davis RW (1977) Screening λgt recombinant clones by hybridization to single plaques in situ. Science 196:180–182
Boston RS, Kodrzycki R, Larkins BA (1986) Transcriptional and posttranscriptional regulation of maize zein genes. In: Shannon LM, Chrispeels MJ (eds) Molecular biology of seed storage proteins and lectins. Ninth symposium in plant physiology. University of California, Riverside, Calif, pp 117–126
Brown JWS, Wandelt C, Feix G, Neuhaus G, Schweiger HG (1986) The upstream regions of zein genes—Sequence analysis and expression in the unicellular alga Acetabularia. Eur J Cell Biol 42:161–170
Carlson DP, Ross J (1984) α-Amanitin-insensitive transcription of mouse β-major-globin 5′-flanking and structural gene sequences correlates with mRNA expression. Proc Natl Acad Sci USA 81:7782–7786
Dale RMK, McClure BA, Houchins JP (1984) A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: Application to sequencing the corn mitochondrial 18S rDNA. Plasmid 13:31–40
Esen A (1986) Separation of alcohol-soluble proteins (zeins) from maize into three fractions by differential solubility. Plant Physiol 80:623–627
Forde BG, Heyworth A, Pywell J, Kreis M (1985) Nucleotide sequence of a B1 hordein gene and the identification of possible upstream regulatory elements in endosperm storage protein genes from barley, wheat and maize. Nucleic Acids Res 13:7327–7339
Geraghty DE, Messing J, Rubenstein I (1982) Sequence analysis and comparison of cDNAs of the zein multigene family. EMBO J 1:1329–1335
Goldsbrough PB, Gelvin SB, Larkins BA (1986) Expression of maize zein genes in transformed sunflower cells. Mol Gen Genet 202:374–381
Grindlay GJ, Lanyon WG, Paul J (1984) Alternative sites of transcription initiation upstream of the canonical cap site in human γ-globin and β-globin genes. Nucleic Acids Res 12:1811–1820
Hagen G, Rubenstein I (1981) Complex organization of zein genes in maize. Gene 13:239–249
Heidecker G, Messing J (1986) Structural analysis of plant genes. Annu Rev Plant Physiol 37:439–466
Hu NT, Peifer MA, Heidecker G, Messing J, Rubenstein I (1982) Primary structure of a genomic zein sequence of maize. EMBO J 1:1337–1342
Kreis M, Shewry PR, Forde BG, Miflin BJ (1985) Structure and evolution of seed storage proteins and their genes with particular reference to those of wheat, barley and rye. Oxford Surv Plant Mol Cell Biol 2:253–327
Langridge P, Feix G (1983) A zein gene of maize is transcribed from two widely separated promoter regions. Cell 34:1015–1022
Langridge P, Pinter-Toro JA, Feix G (1982) Zein precursor mRNAs from maize endosperm. Mol Gen Genet 187:432–438
Langridge P, Brown JWS, Pinter-Toro JA, Feix G (1985) Expression of zein genes in Acetabularia mediterranea. Eur J Cell Biol 39:257–264
Loennen WAM, Blattner FR (1983) Lambda Charon vectors (Ch32, 33, 34 and 35) adapted for DNA cloning in recombinant-deficient hosts. Gene 26:171–179
Maniatis T, Fritsch E, Sambrook J (1982) Molecular cloning. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York pp 80–85
Marks MD, Larkins BA (1982) Analysis of sequence microheterogeniety among zein messenger RNAs. J Biol Chem 260:9976–9983
Marks MD, Lindell JS, Larkins BA (1985a) Nucleotide sequence analysis of zein mRNAs from maize endosperm. J Biol Chem 260:16451–16459
Marks MD, Lindell JS, Larkins BA (1985b) Quantitative analysis of the accumulation of zein mRNA during maize endosperm development. J Biol Chem 260:16445–16450
Pedersen K, Devereux J, Wilson DR, Sheldon E, Larkins BA (1982) Cloning and sequence analysis reveal structural variation among related zein genes in maize. Cell 29:1015–1026
Rigby PWJ, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113:237–251
Rivin CJ, Zimmer EA, Walbot V (1982) Isolation of DNA and DNA recombinants from maize. In: Sheridan WF (ed) Maize for biological research. Plant Molecular Biology Association, Charlottesville, Virginia, pp 155–160
Roussell DR (1986) Identification of 5′ noncoding sequence required for maximal transcription of a zein gene. Ph.D. Dissertation, Purdue University
Slightom JL, Blechel AE, Smithies O (1980) Human fetal G- and A-globin genes: complete nucleotide sequences suggest that DNA can be exchanged between these duplicated genes. Cell 21:627–638
Soave C, Salamini F (1984) Organization and regulation of zein genes in maize endosperm. Philos Trans R Soc Lond [Biol] 304:341–347
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Spena A, Viotti A, Pirrotta V (1982) A homologous repetitive block structure underlies the heterogeneity of heavy and light chain zein genes. EMBO J 1:1589–1594
Spena A, Viotti A, Pirrotta V (1983) Two adjacent genomic zein sequences: Structure, organization and tissue-specific restriction pattern. J Mol Biol 169:799–811
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268
Wahl GM, Stern M, Stark GR (1979) Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethylpaper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci USA 76:3683–3687
Weiher H, Konig M, Gruss P (1983) Multiple point mutations affecting the Simian Virus 40 enhancer. Science 219:626–631
Weinand U, Feix G (1978) Electrophoretic fractionation and translation in vitro of poly (rA)-containing RNA from maize endosperm. Eur J Biochem 92:605–611
Wilson DR, Larkins BA (1984) Zein gene organization in maize and related grasses. J Mol Evol 20:330–340
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M13 phage cloning vectors and host strains: Nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33:103–119
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Kriz, A.L., Boston, R.S. & Larkins, B.A. Structural and transcriptional analysis of DNA sequences flanking genes that encode 19 kilodalton zeins. Mol Gen Genet 207, 90–98 (1987). https://doi.org/10.1007/BF00331495
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DOI: https://doi.org/10.1007/BF00331495