Abstract
cDNA clones of the two non-allelic sucrose synthase (Ss) genes, Ss2 and Sh, have been isolated from λgt11 expression libraries derived from immature kernel poly(A)+ RNA of sh-deletion and Sh/Sh genotypes of maize respectively. Recombinant clones containing the longest Ss2 and Sh cDNA inserts, each of approximately 2.5 kb size, were characterized and comparatively analyzed. Although the Sh cDNA insert expresses as a sucrose synthase-1 (SS1) β-galactosidase fusion protein (∼ 200 kD) in λ lysogens, the Ss2 cDNA failed to form such a chimeric protein and instead showed a ∼ 70 kD SS2 polypeptide. The Ss2 and Sh cDNAs as hybridization probes on RNA blots of immature kernels detected a larger Ss2 transcript (∼ 2900 b) than the Sh transcript (∼ 2750 b). Because SS1 and SS2 protein subunits are known to be of identical size, the significance of difference in transcript size is not apparent. A comparative restriction enzyme mapping of the two cDNA clones and a genomic Ss2 clone show sequence diversity over the entire lengths of Ss2 and Sh clones. Interestingly, restriction endonuclease sites around the 3′ ends are more conserved than the 5′ ends of these two genes. Genetic data indicate that the Ss2 locus is on chromosome 9 and molecular mapping using the Ss2 cDNA clone on recombinant inbred lines and B-A translocations stocks suggest that Ss2 is about 20 map units away from the Wx locus on 9L.
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References
Bachmair A, Finley D, Varshavsky A: In vivo half-life of a protein is a function of its amino-terminal residue. Science 234: 179–186 (1986).
Beckett JB: B-A translocations in maize. J Hered 69: 27–36 (1978).
Burr B, Burr FA: Controlling-element events at the shrunken locus in maize. Genetics 98: 143–156 (1981).
Burr B, Burr FA: Ds controlling elements of maize at the shrunken locus are large and dissimilar insertions. Cell 29: 977–986 (1982).
Chourey PS: Genetic control of sucrose synthetase in maize endosperm. Mol Gen Genet 184: 372–376 (1981).
Chourey PS, Nelson OE: The enzymatic deficiency conditioned by the shrunken-1 mutations in maize. Biochem Genet 14: 1041–1055 (1976).
Chourey PS, Nelson OE: Interallelic complementation at the sh locus in maize at the enzyme level. Genetics 91: 317–325 (1979).
Chourey PS, Still PE: Maize sucrose synthetase gene — A model system for structure function analysis in higher plants. In: Reddy GM, Coe EHJr (eds) Proc. Intl. Symp. on Gene Structure and Function in Higher Plants. Oxford & IBM Publishing Co., New Delhi (1983) pp 207–212.
Chourey PS, Latham MD, Still PE: Expression of two sucrose synthetase genes in endosperm and seedling cells of maize: Evidence of tissue specific polymerization of protomers. Mol Gen Genet 203: 251–255 (1986).
Chourey PS, Still PE, DeRobertis GA: Altered temporal and/or spatial expression of sucrose synthetase genes in McClintock's sh-m6258 and Ac induced revertant allele in maize. Cold Spring Harbor Meetings, p 65 (1986).
Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: version II. Pl Mol Biol Reporter 1: 19–21 (1983).
Dooner HK: Regulation of the enzyme UFGT by the controlling element Ds in bz-m4, an unstable mutant in maize. Cold Spring Harbor Symp Quant Biol 45: 457–462 (1981).
Doring HP, Geiser M, Starlinger P: Transposable element Ds at the shrunken locus in Zea mays. Mol Gen Genet 184: 377–380 (1981).
Echt CS, Chourey PS: A comparison of two sucrose synthetase isozymes from normal and shrunken maize. Pl Physiol 79: 530–536 (1985).
Evola SV, Burr FA, Burr B: The suitability of restriction fragment length polymorphism as genetic markers in maize. Theor Appl Genet 71: 765–771 (1986).
Geiser M, Weck E, Doring HP, Werr W, Courage-Tebbe U, Tillman E, Starlinger P: Genomic clones of a wild-type allele and a transposable element-induced mutant allele of the sucrose synthase gene of Zea mays L. The EMBO J 1: 1455–1460 (1982).
Huynh TV, Young RA, Davis RW: Constructing and screening cDNA libraries in λgt10 and λgt11. In: Glover D (ed.) DNA Cloning, Vol. 1: A Practical Approach. IRL Press, Oxford (1985) pp 49–78.
Johnson DA, Gautsch JW, Sportsman JR, Elder JH: Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techn 1: 3–8 (1984).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227: 680–685 (1970).
Maniatis T, Fritsch EF, Sambrook J: In: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).
McCarty DR, Shaw JR, Hannah LC: The cloning, genetic mapping, and expression of the constitutive sucrose synthase locus of maize. Proc Natl Acad Sci USA 83: 9099–9103 (1986).
McCormick S, Mauvais J, Fedoroff N: Evidence that the two sucrose synthetase genes in maize are related. Mol Gen Genet 187: 494–500 (1982).
Rigby PW, Dieckmann M, Rhodes C, Berg P: Labelling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113: 237–251 (1977).
Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517 (1975).
Springer B, Werr W, Starlinger P, Bennett DC, Zokolica M, Freeling M: The shrunken gene on chromosome 9 of Zea mays L. is expressed in various plant tissues and encodes an anaerobic protein. Mol Gen Genet 205: 461–468 (1986).
Stern DB, Newton KJ: Isolation of intact plant mitochondrial RNA using aurintricarboxylic acid. Pl Mol Biol Reporter 1: 8–15 (1984).
Werr W, Frommer W-B, Maas C, Starlinger P: Structure of the sucrose synthase gene on chromosome 9 of Zea mays L. The EMBO J 4: 1373–1380 (1985).
Young RA, Davis RW: Efficient isolation of genes by using antibody probes. Proc Natl Acad Sci USA 80: 1194–1198 (1983).
Young RA, Davis RW: Yeast RNA polymerase II genes: isolation with antibody probes. Science 222: 778–782 (1983).
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Gupta, M., Chourey, P.S., Burr, B. et al. cDNAs of two non-allelic sucrose synthase genes in maize: cloning, expression, characterization and molecular mapping of the sucrose synthase-2 gene. Plant Mol Biol 10, 215–224 (1988). https://doi.org/10.1007/BF00027398
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DOI: https://doi.org/10.1007/BF00027398