Summary
An EcoRI restriction endonuclease fragment of maize DNA coding for the 19,000 dalton zein protein was cloned in phage λgt WES. The zein gene was identified by the electron microscopic analysis of RNA-DNA hybrids (R-loops) and DNA-DNA hybrids (D-loops). The R-loops were formed with poly(rA)-containing RNA isolated from 18 days post-pollination maize endosperm and showed no intervening non-hybridizing sequences (introns) within their 800 base length. A cDNA clone specific for the 19,000 dalton zein protein formed D-loops in the same position and orientation as the R-loops. The cloned fragment measured 4.4 kilobases (kb), the same size as an EcoRI fragment of maize DNA revealed by “Southern” analysis.
Similar content being viewed by others
References
Benton WD, Davis RW (1977) Screening λgt recombinant clones by hybridization to single plaques in situ. Science 196:180–182
Brack C (1981) DNA electron microscopy. CRC Critical Reviews in Biochemistry (in press)
Burr B, Burr FA (1976) Zein synthesis in maize endosperm by polyribosomes attached to protein bodies. Proc Natl Acad Sci USA 73:515–519
Burr B, Burr FA, Rubinstein I, Simon MN (1978). Purification and translation of zein messenger RNA from maize endosperm protein bodies. Proc Natl Acad Sci USA 75:696–700
Gianazza E, Righetti PG, Pioli F, Galante E, Soave C (1976) Size and charge heterogeneity of zein in normal and opaque-2 maize endosperms. Maydica 21:1–17
Hohn B (1979) In vitro packaging of λ and cosmid DNA. In: Colowick, SP, Kaplan NO (eds) Methods in enzymology 68:299–309
Jones RA, Larkins BA, Tsai CY (1977) Storage protein synthesis in maize III. Developmental changes in membrane-bound polyribosome composition and in vitro protein synthesis of normal and opaque-2 maize. Plant Physiol 59:733–737
Langridge J, Langridge P, Bergquist PL (1980) Extraction of nucleic acids from agarose gels. Anal Biochem 103:264–271
Langridge P, Feix G (1981) (in preparation)
Larkins BA, Hurkmann WJ (1978) Synthesis and deposition of zein in protein bodies of maize endosperm. Plant Physiol 62:256–263
Leder P, Tiemeier D, Enquist L (1977) EK2 derivatives of bacteriophage lambda useful in the cloning of DNA from higher organism: the λgt WES system. Science 196:175–177
Lusky M, Hobom G (1979) Inceptor and origin of DNA replication in lambdoid coliphages I. The λ DNA minimal replication system. Gene 6:137–172
Maniatis T, Hardison RC, Lacy E, Lauer J, O'Connel C, Quon D, Sim GK, Efstratiadis A (1978) The isolation of structural genes from libraries f eukaryotic DNA. Cell 15:687–701
Østergaard E, Paludan K, Hyldig-Nielson J, Jorgensen P, Marcker K (1981). The structure of a chromosomal leghemoglobin gene from soybean. Nature (in press)
Soave C, Suman N, Viotti A, Salamini F (1978) Linkage relationship between regulatory and structural gene loci involved in zein synthesis in maize. Theor Appl Genet 52:263–267
Soberon X, Covarrubias L, Bolivar F (1980) Construction and characterization of new cloning vehicles IV. Deletion derivation of pBR322 and pBR325. Gene 9:287–305
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517
Sternberg N, Tiemeier D, Enquist L (1977) In vitro packaging of a λ dam vector containing EcoRI DNA fragments of E. coli and phage PI. Gene 1:255–280
Sullivan D, Brisson N, Goodehild B, Verma DPS, Thomas DY (1981) Molecular cloning and organization of two leghaemoglobin genomic sequences of soybean. Nature 289:516–518
Sun SM, Slighton JL, Hall TC (1981) Intervening sequences in a plant gene—comparison of the partial sequence of cDNA and genomic DNA of French bean phaseolin. Nature 289:37–41
Viotti A, Sala E, Marotta R, Alberi P, Balducci C, Soave C (1979) Genes and mRNAs coding for zein polypeptides in Zea mays. Eur J Biochem 102:213–222
Wienand U, Feix G (1978) Electrophoretic fractionation and translation in vitro of poly(rA)-containing RNA from maize endosperm. Eur J Biochem 92:605–611
Wienand U, Brüschke C, Feix G (1979) Cloning of double stranded DNAs derived from polysomal mRNA of maize endosperm: isolation and characterization of zein clones. Nucl Acid Res 6:2707–2715
Wienand U, Feix G (1980) Zein specific restriction enzyme fragments of maize DNA. FEBS Lett 116:14–16
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Wienand, U., Langridge, P. & Feix, G. Isolation and characterization of a genomic sequence of maize coding for a zein gene. Molec. Gen. Genet. 182, 440–444 (1981). https://doi.org/10.1007/BF00293933
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00293933