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. 1984 Jul;3(7):1525–1531. doi: 10.1002/j.1460-2075.1984.tb02006.x

Transcription of a zein gene introduced into sunflower using a Ti plasmid vector.

M A Matzke, M Susani, A N Binns, E D Lewis, I Rubenstein, A J Matzke
PMCID: PMC557554  PMID: 6745240

Abstract

A maize genomic clone containing a zein gene (Z4) was inserted into the T-region of the T37 Ti plasmid. Agrobacterium tumefaciens cells carrying this modified Ti plasmid were used to inoculate sunflower stemlets. Callus tissue active in nopaline synthesis was grown from a single transformed cell. DNA analysis of this tissue showed that the zein gene plus T-DNA was present in approximately 12 copies per diploid sunflower genome. A 1000 +/- 100 base RNA homologous to a zein probe could be isolated from the engineered sunflower tissue and the 5' end of this RNA was determined by S1 nuclease mapping. Two transcription start sites were detected. The positions of these transcription start sites and the ratio of the amounts of the two transcripts are identical for the Z4 gene in sunflower and in maize endosperm. Although the zein RNA isolated from the engineered sunflower tissue could be translated in a wheat germ system to yield an immuno-precipitable protein of the expected mol. wt., the presence of the zein protein in the sunflower tissue could not be demonstrated.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barton K. A., Binns A. N., Matzke A. J., Chilton M. D. Regeneration of intact tobacco plants containing full length copies of genetically engineered T-DNA, and transmission of T-DNA to R1 progeny. Cell. 1983 Apr;32(4):1033–1043. doi: 10.1016/0092-8674(83)90288-x. [DOI] [PubMed] [Google Scholar]
  2. Bevan M. W., Chilton M. D. Multiple transcripts of T-DNA detected in nopaline crown gall tumors. J Mol Appl Genet. 1982;1(6):539–546. [PubMed] [Google Scholar]
  3. Bevan M. W., Chilton M. D. T-DNA of the Agrobacterium Ti and Ri plasmids. Annu Rev Genet. 1982;16:357–384. doi: 10.1146/annurev.ge.16.120182.002041. [DOI] [PubMed] [Google Scholar]
  4. Bevan M., Barnes W. M., Chilton M. D. Structure and transcription of the nopaline synthase gene region of T-DNA. Nucleic Acids Res. 1983 Jan 25;11(2):369–385. doi: 10.1093/nar/11.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Binns A. N., Sciaky D., Wood H. N. Variation in hormone autonomy and regenerative potential of cells transformed by strain A66 of Agrobacterium tumefaciens. Cell. 1982 Dec;31(3 Pt 2):605–612. doi: 10.1016/0092-8674(82)90316-6. [DOI] [PubMed] [Google Scholar]
  6. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  7. Caplan A., Herrera-Estrella L., Inzé D., Van Haute E., Van Montagu M., Schell J., Zambryski P. Introduction of genetic material into plant cells. Science. 1983 Nov 18;222(4625):815–821. doi: 10.1126/science.222.4625.815. [DOI] [PubMed] [Google Scholar]
  8. Cochet M., Perrin F., Gannon F., Krust A., Chambon P., McKnight G. S., Lee D. C., Mayo K. E., Palmiter R. Cloning of an almost full-length chicken conalbumin double-stranded cDNA. Nucleic Acids Res. 1979 Jun 11;6(7):2435–2452. doi: 10.1093/nar/6.7.2435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Greve H., Dhaese P., Seurinck J., Lemmers M., Van Montagu M., Schell J. Nucleotide sequence and transcript map of the Agrobacterium tumefaciens Ti plasmid-encoded octopine synthase gene. J Mol Appl Genet. 1982;1(6):499–511. [PubMed] [Google Scholar]
  10. Depicker A., Stachel S., Dhaese P., Zambryski P., Goodman H. M. Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet. 1982;1(6):561–573. [PubMed] [Google Scholar]
  11. Fraley R. T., Rogers S. G., Horsch R. B., Sanders P. R., Flick J. S., Adams S. P., Bittner M. L., Brand L. A., Fink C. L., Fry J. S. Expression of bacterial genes in plant cells. Proc Natl Acad Sci U S A. 1983 Aug;80(15):4803–4807. doi: 10.1073/pnas.80.15.4803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gelvin S. B., Thomashow M. F., McPherson J. C., Gordon M. P., Nester E. W. Sizes and map positions of several plasmid-DNA-encoded transcripts in octopine-type crown gall tumors. Proc Natl Acad Sci U S A. 1982 Jan;79(1):76–80. doi: 10.1073/pnas.79.1.76. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Herrera-Estrella L., Block M. D., Messens E., Hernalsteens J. P., Montagu M. V., Schell J. Chimeric genes as dominant selectable markers in plant cells. EMBO J. 1983;2(6):987–995. doi: 10.1002/j.1460-2075.1983.tb01532.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hu N. T., Peifer M. A., Heidecker G., Messing J., Rubenstein I. Primary structure of a genomic zein sequence of maize. EMBO J. 1982;1(11):1337–1342. doi: 10.1002/j.1460-2075.1982.tb01319.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Langridge P., Feix G. A zein gene of maize is transcribed from two widely separated promoter regions. Cell. 1983 Oct;34(3):1015–1022. doi: 10.1016/0092-8674(83)90559-7. [DOI] [PubMed] [Google Scholar]
  17. Leemans J., Shaw C., Deblaere R., De Greve H., Hernalsteens J. P., Maes M., Van Montagu M., Schell J. Site-specific mutagenesis of Agrobacterium Ti plasmids and transfer of genes to plant cells. J Mol Appl Genet. 1981;1(2):149–164. [PubMed] [Google Scholar]
  18. Lemmers M., De Beuckeleer M., Holsters M., Zambryski P., Depicker A., Hernalsteens J. P., Van Montagu M., Schell J. Internal organization, boundaries and integration of Ti-plasmid DNA in nopaline grown gall tumours. J Mol Biol. 1980 Dec 15;144(3):353–376. doi: 10.1016/0022-2836(80)90095-9. [DOI] [PubMed] [Google Scholar]
  19. Lewis E. D., Hagen G., Mullins J. I., Mascia P. N., Park W. D., Benton W. D., Rubenstein I. Cloned genomic segments of Zea mays homologous to zein mRNAs. Gene. 1981 Aug;14(3):205–215. doi: 10.1016/0378-1119(81)90116-5. [DOI] [PubMed] [Google Scholar]
  20. Murai N., Kemp J. D., Sutton D. W., Murray M. G., Slightom J. L., Merlo D. J., Reichert N. A., Sengupta-Gopalan C., Stock C. A., Barker R. F., Hall T. C. Phaseolin gene from bean is expressed after transfer to sunflower via tumor-inducing plasmid vectors. Science. 1983 Nov 4;222(4623):476–482. doi: 10.1126/science.222.4623.476. [DOI] [PubMed] [Google Scholar]
  21. Park W. D., Lewis E. D., Rubenstein I. Heterogeneity of zein mRNA and protein in maize. Plant Physiol. 1980 Jan;65(1):98–106. doi: 10.1104/pp.65.1.98. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pedersen K., Devereux J., Wilson D. R., Sheldon E., Larkins B. A. Cloning and sequence analysis reveal structural variation among related zein genes in maize. Cell. 1982 Jul;29(3):1015–1026. doi: 10.1016/0092-8674(82)90465-2. [DOI] [PubMed] [Google Scholar]
  23. Ream L. W., Gordon M. P. Crown gall disease and prospects for genetic manipulation of plants. Science. 1982 Nov 26;218(4575):854–859. doi: 10.1126/science.218.4575.854. [DOI] [PubMed] [Google Scholar]
  24. Richter K., Ammerer G., Hartter E., Ruis H. The effect of delta-aminolevulinate on catalase T-messenger RNA levels in delta-aminolevulinate synthase-defective mutants of Saccharomyces cerevisiae. J Biol Chem. 1980 Sep 10;255(17):8019–8022. [PubMed] [Google Scholar]
  25. Roberts B. E., Paterson B. M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330–2334. doi: 10.1073/pnas.70.8.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Shaw C. H., Leemans J., Shaw C. H., van Montagu M., Schell J. A general method for the transfer of cloned genes to plant cells. Gene. 1983 Sep;23(3):315–330. doi: 10.1016/0378-1119(83)90021-5. [DOI] [PubMed] [Google Scholar]
  27. Thomashow M. F., Nutter R., Montoya A. L., Gordon M. P., Nester E. W. Integration and organization of Ti plasmid sequences in crown gall tumors. Cell. 1980 Mar;19(3):729–739. doi: 10.1016/s0092-8674(80)80049-3. [DOI] [PubMed] [Google Scholar]
  28. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Van Montagu M., Schell J. The Ti plasmids of Agrobacterium. Curr Top Microbiol Immunol. 1982;96:237–254. doi: 10.1007/978-3-642-68315-2_13. [DOI] [PubMed] [Google Scholar]
  30. Willmitzer L., Dhaese P., Schreier P. H., Schmalenbach W., Van Montagu M., Schell J. Size, location and polarity of T-DNA-encoded transcripts in nopaline crown gall tumors; common transcripts in octopine and nopaline tumors. Cell. 1983 Apr;32(4):1045–1056. doi: 10.1016/0092-8674(83)90289-1. [DOI] [PubMed] [Google Scholar]
  31. Willmitzer L., Simons G., Schell J. The TL-DNA in octopine crown-gall tumours codes for seven well-defined polyadenylated transcripts. EMBO J. 1982;1(1):139–146. doi: 10.1002/j.1460-2075.1982.tb01137.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zambryski P., Holsters M., Kruger K., Depicker A., Schell J., Van Montagu M., Goodman H. M. Tumor DNA structure in plant cells transformed by A. tumefaciens. Science. 1980 Sep 19;209(4463):1385–1391. doi: 10.1126/science.6251546. [DOI] [PubMed] [Google Scholar]

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