Abstract
High molecular weight DNA released from isolated protoplasts was digested with rare-cutting restriction enzymes and separated by pulsed field gel electrophoresis. The average size of undigested DNA was above 1500 kbp. Digests made with NotI, SfiI, MluI and SalI was hybridized to a probe, common to all genes of theHor2 locus encoding B-hordein polypeptides, and this revealed the maximum size of the locus to be 360 kbp. Two probes, specific for individual B-hordein genes, enabled the identification of two fragment classes in the locus, each containing an equal number of B-hordein genes. Double digests allowed ordering of sites and construction of a map covering 650 kbp around theHor2 locus. No evidence for physical linkage of the two fragment classes was obtained. The possible assignment of the two classes of hybridizing fragments to the B1- and B3-hordein subgroups is discussed.
Article PDF
Similar content being viewed by others
Abbreviations
- bp:
-
basepair
- HMW:
-
High Molecular Weight
- kbp:
-
kilobasepair
- OFAGE:
-
Orthogonal Field Alternation Gel Electrophoresis
- PFGE:
-
Pulsed Field Gel Electrophoresis
- RFLP:
-
Restriction Fragment Length Polymorphism
References
Blake, T.K., S.E. Ullrich &R.A. Nilan: Mapping of theHor3 locus encoding D hordein in barley. Theor. Appl. Genet. 63, 367–371 (1982)
Bojsen, K., D. Abildsten, E.Ø. Jensen, K. Paludan &K.A. Marcker: The chromosomal arrangement of six soybean leghemoglobin genes. EMBO J. 2, 1165–1168 (1983)
Brandt, A., A. Montembault, V. Cameron-Mills &S.K. Rasmussen: Primary structure of a B1 hordein gene from barley. Carlsberg Res. Comm. 50, 333–345 (1985)
Brown, W.R.A.: A physical map of the human pseudoautosomal region. EMBO J. 7, 2377–2385 (1988)
Burke, D.T., G.F. Carle &M.V. Olson: Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science 236, 806–812 (1987)
Burmeister, M., A.P. Monaco, E.F. Gillard, G.J.B. van. Ommen, N.A. Affara, M.A. Ferguson-Smith, L.M. Kunkel &H. Lehrach: A 10-megabase physical map of human Xp21, including the Duchenne muscular dystrophy gene. Genomics 2, 189–202 (1988)
Cameron-Mills, V. &A. Brandt: A γ-hordein gene. Plant Molec. Biol. 4 449–461 (1988)
Carle G.F. &M.V. Olson: Separation of Chromosomal DNA molecules from yeast by orthogonal-field-alternation gel electrophoresis. Nucleic Acids Res. 12, 5647–5664 (1984)
Carle, G.F. &M.V. Olson: An electrophoretic karyotype for yeast. Proc. Nat. Acad. Sci. 82, 3756–3760 (1985)
Daelen, R.A.J. van, J.J. Jonkers &P. Zabel: Preparation of megabase-sized tomato DNA and separation of large restriction fragments by field inversion gel electrophoresis (FIGE). Plant Molec. Biol. 12, 341–352 (1989)
Dean, C., P. van den Elzen, S., Tamaki, M. Black, P. Dunsmuir &J. Bedbrook: Molecular characterization of the rbcS multi-gene family of Petunia (Mitchell). Mol. Gen. Genet. 206 465–474 (1987)
Dunnen, J.T. den, E. Bakker, E.G. Klein Breteler, P.L. Pearson &G.J.B. van Ommen: Direct detection of more than 50% of the Duchenne muscular dystrophy mutations by field inversion gels. Nature 329, 640–642 (1987)
Edwards, G.E., S.P. Robinson, N.J.C. Tyler &D.A. Walker: Photosynthesis by isolated protoplasts, protoplast extracts and chloroplasts of wheat. Plant Physiol. 62, 313–319 (1978)
Faulks, A.J., P.R. Shewry &B.J. Miflin: The polymorphism and structural homology of storage polypeptides (hordein) coded by thehor2 locus in barley (Hordeum vulgare L.). Biochem. Genet. 19, 841–857 (1981)
Feinberg, A.P. &B. Vogelstein: A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13 (1983)
Forde, B.G., A. Heyworth, J. Pywell &M. Kreis: 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 (1985)
Forde, B.G., M. Kreis, M.S. Williamson, R.P. Fry, J. Pywell, P.R. Shewry, N. Bunce &B.J. Miflin: Short tandem repeats shared by B- and C-hordein cDNAs suggest a common origin for two groups of cereal storage protein genes. EMBO J. 4, 9–15 (1985)
Ganal, M.W., N.D. Young &S.D. Tanksley: Pulsed field gel electrophoresis and physical mapping of large DNA fragments in theTm-2a region of chromosome 9 in tomato. Mol. Gen. Genet. 215, 395–400 (1989)
Giese, H. &H.E. Hopp: Influence of nitrogen nutrition on the amount of hordein, protein Z and β-amylase messenger RNA in developing endosperms of barley. Carlsberg Res. Comm. 49, 365–383 (1984)
Guzman, R. &J.R. Ecker: Development of large DNA methods for plants: molecular cloning of Arabidopsis and carrot DNA into yeast. Nucleic Acids Res. 16, 11091–11105 (1988)
Hopp, H.E., S.K. Rasmussen &A. Brandt: Organization and transcription of B1 hordein genes in high lysine mutants of barley. Carlsberg Res. Comm. 48, 201–216 (1983)
Hughes, B.G., F.G. White &M.A. Smith: Effect of plant growth, isolation and purification conditions on barley protoplast yield. Biochem. Physiol. Pflanzen 172, 67–77 (1978)
Jensen, J., J.H. Jørgensen, H.P. Jensen, H. Giese &H. Doll: Linkage of the hordein locihor1 andhor2 with the powdery mildew resistance lociMl-k andMl-a on barley chromosome 5. Theor. Appl. Genet. 58, 27–31 (1980)
Kannangara, C.G., S.P. Gough, B. Hansen, J.N. Rasmussen &D.J. Simpson: A homogenizer with replaceable razor blades for bulk isolation of active barley plastids. Carlsberg Res. Comm. 42, 431–439 (1977)
Kreis, M., P.R. Shewry, B.G. Forde, S. Rahman &B.J. Miflin: Molecular analysis of a mutation conferring the high-lysine phenotype on the grain of barley (Hordeum vulgare). Cell 34, 161–167 (1983)
Lawrance, S.K., C.L. Smith, R. Srivastava, C.R. Cantor &S.M. Weissman: Megabase-scale mapping of the HLA gene complex by pulsed field gel electrophoresis. Science 235, 1387–1390 (1987)
McClelland, M. &M. Nelson: The effect of site specific methylation on restriction endonuclease digestion. Nucleic Acids Res. 13, r201-r207 (1985)
Michiels, E., M. Burmeister &H. Lehrach: Derivation of clones close tomet by preparative field inversion gel electrophoresis. Science 236, 1305–1308 (1987)
Netsvetaev, V.P. &A.A. Sozinov: Linkage studies of genesGleI andHrdF in barley chromosome 5. Barley Genet. Newsl. 12, 13–18 (1982)
Okuno, T. &I. Furusawa: A simple method for the isolation of intact mesophyll protoplasts from cereal plants. Plant Cell Physiol. 18, 1357–1362 (1977)
Poustka, A., T. Pohl, D.P. Barlow, G. Zehetner, A. Craig, F. Michiels, E. Ehrich, A.-M. Frischauf &H. Lehrach: Molecular approaches to mammalian genetics. Cold Spring Harbor Symp. Quant. Biol. 51, 131–139 (1986)
Poustka, A., T.M. Pohl, D.P. Barlow, A.M. Frischauf &H. Lehrach: Construction and use of human chromosome jumping libraries from NotI-digested DNA. Nature 325, 353–355 (1987)
Rahman, S., M. Kreis, B.G. Forde &P.R. Shewry &B.J. Miflin: Hordein-gene expression during development of the barley (Hordeum vulgare) endosperm. Biochem. J. 223, 315–322 (1984)
Rappold, G.A. &H. Lehrach: A long range restriction map of the pseudoautosomal region by partial digest PFGE analysis from the telomere. Nucleic Acids Res. 16, 5361–5377 (1988)
Rasmussen, S.K. &A. Brandt: Nucleotide sequences of cDNA clones for C-hordein polypeptides. Carlsberg Res. Comm. 51, 371–379 (1986)
Rasmussen, S.K., H.E. Hopp &A. Brandt: Nucleotide sequences of cDNA clones for B1-hordein polypeptides. Carlsberg Res. Comm. 48, 187–199 (1983)
Reed, K.C. &D.A. Mann: Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 13, 7207–7221 (1985)
Schwartz, D.C. &C.R. Cantor: Separation of yeast chromosome-sized DNAs by Pulsed Field Gel Electrophoresis. Cell 37, 67–75 (1984)
Shewry, P.R., M. Kreis, S. Parmar, E.J.-L. Lew &D.D. Kasarda: Identification of γ-type hordeins in barley. FEBS Lett. 190, 61–64 (1985)
Shewry, P.R., A.J. Faulks, R.A. Pickering, L.T. Jones, R.A. Finch &B.J. Miflin: The genetic analysis of barley storage proteins. Heredity 44, 383–389 (1980)
Shewry, P.R. &S. Parmar: TheHrdF(Hor5) locus encodes γ-type hordeins. Barley Genet. Newsl. 17, 32–35 (1987)
Smith, C.L., T. Matsumoto, O. Niwa, S. Klco, J.B. Fan, M. Yanagida &C.R. Cantor: An electrophoretic karyotype for Schizosaccharomyces pombe by pulsed field gel electrophoresis. Nucleic Acids Res. 15, 4481–4489 (1987)
Sørensen, M.B., V. Cameron-Mills &A. Brandt: Transcriptional and post-transcriptional regulation of gene expression in developing barley endosperm. Mol. Gen. Genet. 217, 195–201 (1989)
Southern, E.M., R. Anand, W.R.A. Brown &D.S. Fletcher: A model for the separation of large DNA molecules by crossed field gel electrophoresis. Nucleic Acids Res. 15, 5925 (1987)
Sugita, M., T. Manzara, E. Pichersky, A. Cashmore &W. Gruissem: Genomic organization, sequence analysis and expression of all five genes encoding the small subunit of ribulose-1,5-bis-phosphate carboxylase/oxygenase from tomato. Mol. Gen. Genet. 209, 247–256 (1987)
Young, N.D., D. Zamir, M.W. Ganal &S.D. Tanksley: Use of isogenic lines and simultaneous probing to identify DNA markers tightly linked to theTm-2a gene in tomato. Genetics 120, 579–585 (1988)
Author information
Authors and Affiliations
Additional information
Accepted byE. Lund
Rights and permissions
About this article
Cite this article
Sørensen, M.B. Mapping of theHor2 locus in barley by pulsed field gel electrophoresis. Carlsberg Res. Commun. 54, 109 (1989). https://doi.org/10.1007/BF02908303
DOI: https://doi.org/10.1007/BF02908303