Hostname: page-component-8448b6f56d-dnltx Total loading time: 0 Render date: 2024-04-25T01:34:17.539Z Has data issue: false hasContentIssue false

Isolation and characterization of an expressed napin gene from Brassica rapa

Published online by Cambridge University Press:  19 September 2008

Jean C. Kridl*
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
David W. McCarter
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
Ronald E. Rose
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
Donna E. Scherer
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
Deborah S. Knutzon
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
Sharon E. Radke
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
Vic C. Knauf
Affiliation:
Calgene Inc., 1920 Fifth Street, Davis, CA 95616, USA
*
* Correspondence

Abstract

An expressed napin storage protein gene from Brassica rapa, BcNA1, has been cloned and sequenced. The gene is a member of a family of four to seven napin genes in B. rapa and is highly expressed in developing seeds. An expression cassette containing the DNA flanking the napin coding region of BcNA1 has been engineered and demonstrated to function appropriately, as compared with the gene's endogenous expression, in transgenic rapeseed using the β-glucuronidase reporter gene. The B. rapa BcNA1 gene and a B. napus napin gene, gNa, share extremely high nucleotide homology not only throughout their coding regions, but over a DNA locus comprising 4.3 kb. We suggest the gNa gene was contributed by the original B. rapa progenitor of the amphidiploid B. napus.

Type
Research Papers
Copyright
Copyright © Cambridge University Press 1991

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

1

Current address: Vollum L-474, Oregon Health Sciences University, 3181 S.W. Sam Jackson Park Road, Portland, OR 97201, USA

References

Adelman, J.P., Hayflick, J.S., Vasser, M. and Seeburg, P.H. (1983) In vitro deletional mutagenesis for bacterial production of the 20 000-dalton formof human pituitary growth hormone. DNA 2, 183193.CrossRefGoogle Scholar
Allen, R.D., Cohen, E.A., Vonder Haar, R.A., Adams, C.A., Ma, D.P., Nessler, C.L. and Thomas, T.L. (1987) Sequence and expression of a gene encoding an albumin storage protein in sunflower. Molecular and General Genetics 210, 211218.CrossRefGoogle ScholarPubMed
Baszczynski, C.L. and Fallis, L. (1990) Isolation and nucleotidesequence of a genomic clone encoding a new Brassica napus napin gene. Plant Molecular Biology 14, 633635.CrossRefGoogle Scholar
Bernatzky, R. and Tanksley, S.D. (1986) Genetics of actinrelatedsequences in tomato. Theoretical and Applied Genetics 72, 314321.CrossRefGoogle ScholarPubMed
Bhatty, R.S., McKenzie, S.L. and Finlayson, A.J. (1968) The proteins of rapeseed (Brassica napus L) soluble in salt solutions. Canadian Journal of Biochemistry 46, 11911197.CrossRefGoogle ScholarPubMed
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.CrossRefGoogle ScholarPubMed
Buckley, K.J. (1985) Regulation and expression of PhiX174 lysis gene. PhD thesis. University of California at San Diego.Google Scholar
Crouch, M.L., Tenbarge, K.M., Simon, A.E. and Ferl, R. (1983) cDNA clones for Brassica napus seed storage proteins: evidence from nucleotide sequenceanalysis that both subunits of napin are cleaved from a precursor polypeptide. Journal of Molecular and Applied Genetics 2, 273283.Google Scholar
Ericson, M.L., Rodin, J., Lenman, M., Glimelius, K., Josefsson, L-G and Rask, L., (1986) Structure of the 1.7S storage protein, napin, and its precursor. Journal of Biological Chemistry 261, 1457614581.CrossRefGoogle Scholar
Finlayson, A.J., Bhatty, R.S. and Christ, C.M. (1969) Species and varietal differences in the proteins of rapeseed. Canadian Journal of Botany 47, 679685.CrossRefGoogle Scholar
Gatehouse, J.A., Brown, D., Evans, J.M., Gatehouse, L.N., Jobes, D., Preston, P. and Croy, R.R.D. (1987) Sequence of the seed lectin gene from pea (Pisum sativum L.). Nucleic Acids Research 15, 7642.CrossRefGoogle ScholarPubMed
Goldberg, R.B., Barker, S.J. and Perez-Grau, L. (1989) Regulation of gene expression during plant embryogenesis Cell 56, 149160.Google Scholar
Hanahan, D. (1983) Studies on transformation of Escherichia coli with plasmids. Journal of Molecular Biology 166, 557580.CrossRefGoogle ScholarPubMed
Houck, C.M., Shintani, D.K. and Knauf, V.C. (1990) Agrobacterium as a gene transfer agent for plants. Frontiers in Applied Microbiology 4, 117.Google Scholar
Hood, E.H., Helmer, G.L., Fraley, R.T. and Chilton, M-D. (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. Journal of Bacteriology 168, 12911301.CrossRefGoogle Scholar
Jefferson, R.A. (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Molecular Biology Reporter 5, 387405.CrossRefGoogle Scholar
Josefsson, L-G., Lenman, M., Ericson, M.L. and Rask, L. (1987) Structure of a gene encoding the 1.7 S storage protein, napin, from Brassica napus. Journal of Biological Chemistry 262,1219612201.CrossRefGoogle ScholarPubMed
Krebbers, E., Herdies, L., De Clercq, A., Seurinck, J., Leemans, J., Van Damme, J., Segura, M., Gheysen, G., Van Montagu, M. and Vandekerckhove, J. (1988)Determination of the processing sites of an Arabidopsis 2S albumin and characterization of the complete gene family. Plant Physiology 87, 859866.CrossRefGoogle ScholarPubMed
Koning, A., Jones, A., Fillatti, J.J., Comai, L. and Lassner, M.W. (1991) Arrest of embryo development in Brassica napus mediated by modified Pseudomonas aeruginosa exotoxin A. Plant Molecular Biology, in press.CrossRefGoogle Scholar
Lee, W.S., Tzen, J.T.C., Kridl, J.C., Radke, S.E. and Huang, A.H.C. (1991) Maize oleosin is correctly targeted to seed oil bodies in Brassica napus transformed with the maize oleosin gene. Proceedings of the National Academy of Sciences USA 88, 61816185.CrossRefGoogle ScholarPubMed
Loenen, W.A.M. and Blattner, F.R. (1983) Lambda Charon vectors (Ch32, 33, 34 and 35) adapted for DNA cloning in recombinant-deficient hosts.Gene 26, 171179.CrossRefGoogle Scholar
Lonnerdal, B. and Janson, J-C. (1972) Studies on Brassica seed proteins. I. The low molecular weight proteins in rapeseed. Isolation and characterization. Biochimica Biophysica Acta 278, 175183.Google ScholarPubMed
Maniatis, T., Fritsch, E.F. and Sambrook, J. (1982) Molecular Cloning, A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY.Google Scholar
Norrander, J., Kempe, T. and Messing, J. (1983) Construction ofimproved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26, 101106.CrossRefGoogle ScholarPubMed
Radke, S.E., Andrews, B.M., Moloney, M.M., Crouch, M.L., Kridl, J.C. and Knauf, V.C. (1988) Transformation of Brassica napus L. using Agrobacterium tumefaciens: developmentally regulated expression of a reintroduced napin gene. Theoretical and Applied Genetics 75, 685694.CrossRefGoogle Scholar
Radke, S.E., Turner, J.C., Facciotti, D. and Knauf, V.C. (1990) Expression of chimeric genes in developing embryos derived from isolated microspores fromtransgenic Brassica napus plants. pp 410415 in, Nijkamp, H.J.J., Van DerPlas, L.H.W., Van Aartrijk, J.(Eds)Progress in Plant Cellular and Molecular Biology. Dordrecht, The Netherlands, Kluwer Academic.CrossRefGoogle Scholar
Raynal, M., Depigny, D., Grellet, F. and Delseny, M. (1991) Characterization and evolution of napin-encoding genes in radish and related crucifers. Gene 99, 7786.CrossRefGoogle ScholarPubMed
Rose, R.E., DeJesus, C.E., Moylan, S.L., Ridge, N.P., Scherer, D.E. and Knauf, V.C. (1987) The nucleotide sequence of a cDNA clone encoding acyl carrier protein (ACP) from Brassica rapa seeds. Nucleic Acids Research 15,7197.CrossRefGoogle Scholar
Sanger, F., Nicklen, S. and Coulson, A.R. (1977) DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences USA 74, 54635467.CrossRefGoogle ScholarPubMed
Scherer, D.E. and Knauf, V.C. (1987) Isolation of a cDNA clone for the acyl carrier protein-I of spinach. Plant Molecular Biology 9,127134.CrossRefGoogle ScholarPubMed
Scofield, S.R. and Crouch, M.L. (1987) Nucleotide sequence of a member of the napin storage protein family from Brassica napus.Journal of Biological Chemistry 262, 1220212208.CrossRefGoogle ScholarPubMed
U, N. (1935) Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization. Japanese Journal of Botany 7, 389452.Google Scholar
Vieira, J. and Messing, J. (1987) Vectors for cloning DNA: production of single-stranded plasmid DNA. Methods in Enzymology 153, 311.CrossRefGoogle Scholar
Vodkin, L.O., Rhodes, P.R. and Goldberg, R.B. (1983) A lectin gene insertion has the structural features of a transposable element. Cell 34, 10231031.CrossRefGoogle ScholarPubMed