Summary
A cDNA library was made from poly(A+) RNA isolated from developing oat seeds, and oat globulin cDNA clones were identified by hybridization with synthetic oligonucleotides. Globulin clones were characterized by restriction enzyme mapping and cross-hybridization analysis. Based on these comparisons, four classes of globulin clones were distinguished. These clones hybridized to multiple DNA fragments in restriction enzyme digests of oat genomic DNA, indicating that the genes exist in a multigene family. The nucleotide sequence of one of the globulin cDNA clones was determined. The amino acid sequence derived from the DNA sequence verified its identity as an oat globulin and confirmed that the protein is synthesized as a precursor similar to legume 11S storage globulins. The basic polypeptide encoded at the 3′ end of the mRNA was found to be homologous to the basic polypeptides of other 11S seed globulins.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ds:
-
double stranded
- kb:
-
kilobase
References
Adeli K, Altosaar I: Role of endoplasmic reticulum in biosynthesis of oat globulin precursors. Plant Physiol 73:949–955, 1983.
Argos P, Narayana SVL, Nielsen NC: Structural similarity between legumin and vicilin storage proteins from legumes, EMBO J (in press).
Bailey JM, Davidson N: Methyl mercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem 70:75–85, 1976.
Brinegar AC, Peterson DM: Separation and characterization of oat globulin polypeptides Arch Biochem Biophys 219:71–79, 1982.
Brinegar AC, Peterson DM: Synthesis of oat globulin precursors Plant Physiol 70:1767–1769, 1982.
Buell GN, Wickens MP, Payvar F, Schimke RT: Synthesis of full length cDNAs from four partially purified oviduct mRNAs J Biol Chem 253:2471–2482, 1978.
Croy RRD, Lycett GW, Gatehouse JA, Yarwood JN, Boulter D: Cloning and analysis of cDNAs encoding plant storage protein precursors Nature 295:76–79, 1982.
Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: Version II. Plant Mol Biol Reporter 1:19–21, 1983.
Derbyshire E, Wright DJ, Boulter D: Legumin and vicilin, storage proteins of legume seeds. Phytochemistry 15:3–24, 1976.
Domoney C, Casey R: Measurement of gene number for seed storage proteins in Pisum. Nucleic Acids Res 13:687–699, 1985.
Gough NM, Gough J, Metcalf D, Kelso A, Nicola NA, Burgess AW, Dunn AR: Molecular cloning of cDNA encoding a murine haematopoetic growth regulator, granulocyte-macrophage colong stimulating factor. Nature 309:763–767, 1984.
Kafatos FC, Jones CW, Efstratiadis A: Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure, Nucleic Acids Res 7:1541–1552, 1979.
Krayev AS, Kramerow DA, Skryabim KG, Ryslov AP, Bayer AA, Georgien CP: The nucleotide sequence of the ubiquitous repetitive DNA sequence B1 complementary to the most abundant class of mouse fold-back RNA, Nucleic Acids Res 8:1201–1215, 1980.
Lycett GW, Croy RRD, Shirgat AH, Boulter D: The complete nucleotide sequence of a legumin gene from pea (Pisum sativum L). Nucleic Acids Res 12:4493–4506, 1984.
Lycett GW, Delauney AJ, Zhao W, Gatehouse JA, Croy RRD, Boulter D: Two cDNA clones coding for the legumin protein of Pisum sativum L. contain sequence repeats. Plant Mol Biol 3:91–96, 1984.
Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A laboratory Manual. Cold Spring Harbor Laboratory, New York, 1982.
Marco YA, Thanh VH, Tumer NE, Scallon BJ, Nielsen NC: Cloning and structural analysis of DNA complementary to A2B1a subunit of glycinin. J Biol Chem 259:13436–13441, 1984.
Matlashewski GJ, Adeli K, Altosaar I, Shewry PR, Miflin BJ: In vitro synthesis of oat globulin. FEBS Lett 145:2085–2092, 1982.
Matteucci MD, Caruthers MH: The synthesis of oligodeoxypyrimidines on a polystyrene support. Tetrahedron Lett. 21:719–722, 1980.
Maxam A, Gilbert W: Sequencing DNA with base-specific chaeical cleavages. Methods Enzymol. 6S:499–559, 1980.
Peterson DM: Subunit structure and composition of oat seed globulin. Plant Physiol 62:506–509, 1978.
Rossi HA, Luthe DS: Isolation and characterization of oat globulin messenger RNA. Plant Physiol 72:578–582, 1983.
Saigo RH, Peterson DM, Holy J: Development of protein bodies in oat starch endosperm, Can J Bot 61:1206–1215, 1983.
Scallon BJ, Thanh VH, Floener LA, Nielsen NC: Identification and characterization of DNA clones encoding Group-II glycinin subunits. Theor Appl Genet (submitted) 1985.
Simon AE, Tenbarge KM, Scotfield SR, Finkelstein RR, Crouch ML: Nucleotide sequence of a cDNA clone of Brassica napus 12S storage protein shows homology with legumin from Pisum sativum. Plant Mol Biol (submitted) 1985.
Southern EM: Detection of specific DNA sequences among DNA fragments separated by electrophoresis, J Mol Biol 98:503–517, 1975.
Walburg G, Larkins BA: Oat Seed Globulin. Subunit characterization and demonstration of its synthesis as a precursor. Plant Physiol 72:161–165, 1983.
Wickens MP, Buell GN, Schimke RT: Synthesis of double-stranded DNA complementary to lysozyme, ovomucoid and ovalbumin mRNAs. J Biol Chem 253:2483–2495, 1978.
Author information
Authors and Affiliations
Additional information
Author to whom correspondence should be addressed. Journal paper number 10460 of the Purdue Agricultural Experimental Station.
Rights and permissions
About this article
Cite this article
Walburg, G., Larkins, B.A. Isolation and characterization of cDNAs encoding oat 12S globulin mRNAs. Plant Mol Biol 6, 161–169 (1986). https://doi.org/10.1007/BF00021485
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00021485