Abstract
The xylanase gene from Cryptococcus albidus contains seven introns. Genomic and cDNA clones under the control of the CaMV 35S promoter were transferred into tobacco plants using Agrobacterium-mediated cell transformation. The genes were transcribed and the mRNAs were amplified by the polymerase chain reaction using primers on each side of the intron region. About 90% of the amplification products from plants transformed with the genomic clone corresponded to the size of the pre-mRNA (1.2 kb) and 10% represented the spliced product (0.85 kb). The 0.85 kb fragment was cloned and sequenced and the result indicated that the introns from the xylanase gene were accurately spliced by the plant cells.
Similar content being viewed by others
References
Baulcombe DC, Saunders GR, Bevan MW, Mayo MA, Harrison BD: Expression of biologically active viral satellite RNA from the nuclear genome of transformed plants. Nature 321: 446–449 (1986).
Barta A, Sommergruber K, Thompson D, Hartmuth K, Matzke MA, Matzke AJM: The expression of a nopaline synthase-human growth hormone chimaeric gene in transformed tobacco and sunflower tissue. Plant Mol Biol 6: 347–357 (1986).
Beggs JD, van den Berg J, van Ooyen A, Weissmann C: Abnormal expression of chromosomal rabbit β-globin gene in Saccharomyces cerevisiae. Nature 283: 835–840 (1980).
Bevan MW: Binary Agrobacterium vectors for plant transformation. Nucl Acids Res 12: 8711–8721 (1984).
Boucher F, Morosoli R, Durand S: Complete nucleotide sequence of the xylanase gene from the yeast Cryptococcus albidus. Nucl Acids Res 16: 9874 (1988).
Brown JWS: A catalogue of splice junction and putative point sequences from plant introns. Nucl Acids Res 14: 9549–9559 (1986).
Brown JWS, Feix G, Frendewey D: Accurate in vitro splicing of two pre-mRNA plant introns in a HeLA cell nuclear extract. EMBO J 5: 2749–2758 (1986).
Ditta G, Stanfield S, Corbin D, Helinski DR: Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77: 7347–7351 (1980).
Goodall GJ, Filipowitz W: The AU-rich sequences present in the introns of plant nuclear pre-mRNAs are required for splicing. Cell 58: 473–483 (1989).
Goodall GJ, Filipowitz W: The minimum functional length of pre-mRNA intron in monocots and dicots. Plant Mol Biol 14: 727–733 (1990).
Hanley BA, Schuler MA: Plant intron sequences: evidence for distinct group of introns. Nucl Acids Res 16: 7159–7176 (1988).
Hartmuth K, Barta A: In vitro processing of a plant pre-mRNA in HeLa cell nuclear extract. Nucl Acids Res 14: 7513–7528 (1986).
Hunt AG, Mogen BD, Chu NM, Chua N-M: The SV40 small intron is accurately and efficiently spliced in tobacco cells. Plant Mol Biol 16: 375–379 (1991).
Horsch RB, Fry JE, Hoffman NL, Wallroth M, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).
Langford CJ, Klinz F-J, Donath C, Gallwitz D: Point mutations identify the conserved, intron-contained TAC-TAAC box as an essential splicing signal sequence in yeast. Cell 36: 645–653 (1984).
Moreau A, Durand S, Morosoli R: Expression of xylanase from Cryptococcus albidus in Saccharomyces cerevisiae. Submitted to Genes.
Ruskin B, Pikielny CW, Rosbash M, Green MR: Alternative branch points are selected during splicing of a yeast pre-mRNA in mammalian and yeast extracts. Proc Natl Acad Sci USA 83: 2022–2026 (1986).
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1987).
Van Santen VL, Spritz RA: Splicing of plant pre-mRNAs in animal systems and vice versa. Gene 56: 253–265 (1987).
Verwoerd TC, Dekker BMM, Hoekema A: A small-scale procedure for the rapid isolation of plant RNAs. Nucl Acids Res 17: 2362 (1989).
Wiebauer K, Herrero J-J, Filipowicz W: Nuclear pre-mRNA processing in plants: distinct modes of 3′-splicesite selection in plants and animals. Mol Cell Biol 8: 2042–2051 (1988).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Laliberté, J.F., Nicolas, O., Durand, S. et al. The xylanase introns from Cryptococcus albidus are accurately spliced in transgenic tobacco plants. Plant Mol Biol 18, 447–451 (1992). https://doi.org/10.1007/BF00040660
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00040660