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. 1987 Aug;84(4):1185–1192. doi: 10.1104/pp.84.4.1185

Factors Affecting the Level of Kanamycin Resistance in Transformed Sunflower Cells

Robert Nutter 1,1, Nicholas Everett 1, Dorothy Pierce 1, Lucy Panganiban 1, Patricia Okubara 1, Rosanna Lachmansingh 1, Desmond Mascarenhas 1, Heather Welch 1, Irvin Mettler 1, Lucille Pomeroy 1, Jill Johnson 1, John Howard 1
PMCID: PMC1056749  PMID: 16665582

Abstract

A 230 base pair DNA segment containing the sequences 5′ to the 700 to 750 nucleotide (nt) transcript 7′ (ORF 3; RF Barker, KB Idler, DV Thompson, JD Kemp 1983 Plant Mol Biol 2: 335-350) of the octopine tumor inducing plasmid pTiA6 has been isolated. This region has (a) 180 base pairs of DNA upstream of the TATA box, (b) the start of RNA synthesis, and (c) the entire 5′ untranslated region of the gene. We have fused this presumed promoter fragment to the neomycin phosphotransferase II (NPTII) gene from Tn5 in a plant expression cassette. After recombination into a tumor inducing plasmid delivery plasmid, this cassette confers selectable kanamycin resistance to transformed sunflower cells. Removal of the out-of-frame ATG in the 5′ leader sequence of the NPTII gene by two different modifications increased both the levels of NPTII enzyme activity and the ID50 for kanamycin in the tumor cells. The promoter region of the transcript 7 gene gives levels of kanamycin resistance equivalent to the nopaline synthase promoter and octopine synthase promoter when used in the same constructions and assayed in the same tissues.

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Selected References

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

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