Open AccessIntegration and expression of a rabbit liver cytochrome P-450 gene in transgenic Nicotiana tabacum.
Author(s) -
Kazuki Saito,
Masaaki Noji,
Shigeru Ohmori,
Yumiko Imai,
Isamu Murakoshi
Publication year - 1991
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.88.16.7041
Subject(s) - nicotiana tabacum , biology , agrobacterium tumefaciens , microbiology and biotechnology , cytochrome , agrobacterium , transgene , ti plasmid , transformation (genetics) , cytochrome p450 , plasmid , gene , biochemistry , metabolism , enzyme
Cytochrome P-450 is involved in the oxidative metabolism of a broad range of substrates. We have made a chimeric construct, pSN002, containing the cDNA for rabbit liver cytochrome P-450 (IIC14) under the control of the TR2' promoter for mannopine synthase in the Agrobacterium Ti plasmid. Nicotiana tabacum was transformed with Agrobacterium tumefaciens harboring a cointegrated plasmid pSN002::pGV2260. The presence of mRNA and of the translated protein from the chimeric cytochrome P-450 gene in transgenic plants was confirmed by RNA blot hybridization and by Western blot and immunohistochemical analyses, respectively. The transformants in which the foreign cytochrome P-450 protein is expressed show marked phenotypic changes, notably a tendency rapidly to senesce. We detected 2-propenylpyrrolidine, a degradative metabolite of nicotine alkaloids, in transgenic tobacco showing this pronounced phenotypic change. Such metabolism is likely to be due to the effect of senescence and not directly to the presence of the cytochrome P-450.