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Characterization and Expression of a Wound-Inducible Proteinase Inhibitor II Gene from Potato

  • Robert W. Thornburg
  • Gyn An
  • Thomas A. Cleveland
  • Clarence A. Ryan
Part of the Basic Life Sciences book series (BLSC, volume 41)

Summary

An 8-kb fragment of potato DNA containing the potato Inhibitor II gene (called Inhibitor IIK gene) was isolated from a library of EcoRI-restricted potato DNA in λ bacteriophage. A 2.6-kb TaqI fragment containing the gene was subcloned into the plasmid pUC13 and sequenced. This gene is one of 10 to 12 Inhibitor II genes identified by analysis of fragments from restriction enzyme digests of total Russet Burbank genomic DNA by hybridization with nick-translated tomato Inhibitor II complementary DNA (cDNA). A 1.0-kb fragment from the 3’ region of the gene, containing only 11 bp of the open reading frame, hybridized strongly with mRNA species from wounded leaves, but only weakly with mRNA from unwounded leaves or tubers. This indicated that the gene is a wound-inducible gene. Approximately 900 bases from the 5’ region of the gene and 1,000 bases from the 3’ region were fused with the open reading frame of the reporter gene encoding the enzyme chloramphenicol acetyl transferase (CAT) gene. The chimeric gene was cloned into a binary transformation vector derived from the Ti plasmid and successfully used to transform tobacco plants with a wound-inducible CAT gene. Thus, nucleotide sequences in the 5’ and 3’ regions of the Inhibitor IIK gene are sufficient for wound-inducible expression of CAT activity. These experiments also demonstrate that biochemical components present in cells of wounded tobacco can recognize the potato Inhibitor IIK regulation sequences and can activate the expression of the CAT gene.

Keywords

Chloramphenicol Acetyl Transferase Wounded Leaf Transform Tobacco Plant Chloramphenicol Acetyl Transferase Activity Potato Inhibitor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Robert W. Thornburg
    • 2
  • Gyn An
    • 1
  • Thomas A. Cleveland
    • 3
  • Clarence A. Ryan
    • 1
  1. 1.Institute of Biological ChemistryWashington State UniversityPullmanUSA
  2. 2.Department of Biochemistry and BiophysicsIowa State UniversityAmesUSA
  3. 3.U.S. Department of Agriculture, Agricultural Research ServiceSouthern Regional Research LaboratoryNew OrleansUSA

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