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Molecular cloning and characterization of multiple isoforms of the snowdrop (Galanthus nivalis L.) lectin

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Abstract

Screening of a copy-DNA (cDNA) library constructed from RNA isolated from young developing ovaries of snowdrop (Galanthus nivalis) resulted in the isolation of five lectin clones which clearly differed from each other with regard to their nucleotide sequence and deduced amino-acid sequence. Sequence comparison between the coding regions of different lectin cDNAs revealed the highest homology between lectin clones LECGNA 3 and LECGNA 5, showing 96.4% and 93.6% similarity at the nucleotide level and at the deduced amino-acid level, respectively, whereas lectin clones LECGNA 1 and LECGNA 3 showed the lowest homology of 81.6% and 68.6% for the nucleotide sequence and the amino-acid sequence, respectively. Only very few lectin cDNA clones containing a polyadenylated tail could be isolated. Moreover all these cDNA clones were derived from isolectin 3 and showed some variability within the length of the 3′ untranslated region. The major transcription initiation site was located 30 bases upstream from the AUG codon as could be deduced from primer-extension analysis. Taking into account the small 5′ untranslated region of the lectin clones, the size of the lectin mRNA, which is approx. 780 nucleotides as determined by Northern blot analysis, is in good agreement with the length of the cDNA clones isolated. Besides the ovary tissue, both the leaf and the flower tissue were also shown to express the lectin mRNA in a flowering snowdrop plant.

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Abbreviations

cDNA:

copy DNA

kb:

kilobase (standardsymbol)

References

  • Balzarini, J., Schols, D., Neyts, J., Van Damme, E., Peumans, W., De Clercq, E. (1991) The replication of human immunodefficiency virus and cytomegalovirus is markedly inhibited by α-(1–3)and α-(1–6)-D-mannose-specific plant lectins. Antimicrob. Agents Chemother. 35, 410–416

    Google Scholar 

  • de Kochko, A., Hamon, S. (1990) A rapid and efficient method for the isolation of restrictable total DNA from plants of the genus Abelmoschus. Plant Mol. Biol. Rep. 8, 3–7

    Google Scholar 

  • Dellaporta, S.L., Wood, J., Hicks, J.B. (1983) A plant DNA minipreparation: version II. Plant Mol. Biol. Rep. 1, 19–21

    Google Scholar 

  • Denhardt, D.J., Dressler, D., Ray, D.S. (1978) The single-stranded DNA phages. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Feldsted, R.L., Egorin, M.J., Leavitt, R.D., Bachur, N.R. (1977) Recombination of subunits of Phaseolus vulgaris isolectins. J. Biol. Chem. 252, 2967–2971

    Google Scholar 

  • Finkelstein, R.R., Crouch, M.L. (1986) Rapeseed embryo development in culture on high osmoticum is similar to that in seeds. Plant Physiol. 81, 907–912

    Google Scholar 

  • Joshi, C.P. (1987) Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucleic Acids Res. 15, 9627–9640

    Google Scholar 

  • Maniatis, T., Fritsch, E.F., Sambrook, J. (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Mierendorf, R.C., Pfeffer, D. (1987) Direct sequencing of denatured plasmid DNA. Methods Enzymol. 152, 556–562

    Google Scholar 

  • Peumans, W.J., Stinissen, H.M., Carlier, A.R. (1982) A genetic basis for the origin of six different isolectins in hexaploid wheat. Planta 154, 561–567

    Google Scholar 

  • Sanger, F., Nicklen, S., Coulson, A.R. (1977) DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463–5467

    Google Scholar 

  • Shibuya, N., Goldstein, I.J., Van Damme, E.J.M., Peumans, W.J. (1988a) Binding properties of a mannose-specific lectin from the snowdrop (Galanthus nivalis) bulb. J. Biol. Chem. 263, 728–734

    Google Scholar 

  • Shibuya, N., Berry, J.E., Goldstein, I.J. (1988b) One-step purification of murine IgM and human α2-macroglobulin by affinity chromatography on immobilized snowdrop bulb lectin. Arch. Biochem. Biophys. 267, 676–680

    Google Scholar 

  • Siflow, C.D., Hammett, J.R., Key, J.L. (1979) Sequence complexity of polyadenylated ribonucleic acid from soybean suspension culture cells. Biochemistry 18, 2725–2731

    Google Scholar 

  • Smith, J.J., Raikhel, N.V. (1989) Nucleotide sequences of cDNA clones encoding wheat germ agglutinin isolectins A and D. Plant Mol. Biol. 13, 601–603

    Google Scholar 

  • Spiro, R.G. (1970) Glycoproteins. Annu. Rev. Biochem. 39, 599–638

    Google Scholar 

  • Van Damme, E.J.M., Peumans, W.J. (1990a) Developmental changes and tissue distribution of lectin in Galanthus nivalis and Narcissus cv. Carlton. Planta 182, 605–609

    Google Scholar 

  • Van Damme, E.J.M., Peumans, W.J. (1990b) Isolectins in Narcissus: complexity, inter and intraspecies differences and developmental control. Physiol. Plant. 79, 1–6

    Google Scholar 

  • Van Damme, E.J.M., Allen, A.K., Peumans, W.J. (1987) Isolation and characterization of a lectin with exclusive specificity towards mannose from snowdrop (Galanthus nivalis) bulbs. FEBS Lett. 215, 140–144

    Google Scholar 

  • Van Damme, E.J.M., Allen, A.K., Peumans, W.J. (1988) Related mannose specific lectins from different species of the family Amaryllidaceae. Physiol. Plant. 73, 52–57

    Google Scholar 

  • Van Damme, E.J.M., Goldstein, I.J., Peumans, W.J. (1991a) Comparative study of related mannose-binding lectins from Amaryllidaceae and Alliaceae species. Phytochemistry 30, 509–514

    Google Scholar 

  • Van Damme, E.J.M., Kaku, H., Perini, F., Goldstein, I.J., Peeters, B., Yagi, F., Decock, B., Peumans, W.J. (1991b) Biosynthesis, primary structure and molecular cloning of snowdrop (Galanthus nivalis L.) lectin. Eur. J. Biochem., in press

  • Williams, J.G., Mason, P.J. (1985) Hybridisation in the analysis of RNA. In: Nucleic acid hybridisation, a practical approach. Hames, B.D., Higgins, S.J., eds. IRL Press, Oxford

    Google Scholar 

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This work was supported in part by grants of the ‘Nationale Bank’ and the National Fund for Scientific Research (Belgium) (F.G.W.O. grant 20059.89 N). W.P. is Research Director and E.V.D.T., Research Assistant of this fund. We wish to thank V. Feytons and R. De Greef for expert help with DNA sequencing, Professor F. Van Leuven for helping with computer analyses, and Professor W. Rombauts for his hospitality during the stay in his laboratory.

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Van Damme, E.J.M., De Clercq, N., Claessens, F. et al. Molecular cloning and characterization of multiple isoforms of the snowdrop (Galanthus nivalis L.) lectin. Planta 186, 35–43 (1991). https://doi.org/10.1007/BF00201495

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