Expression of functional mammalian P450 2E1 in hairy root cultures
| Autor: | Angela M. Wilson, Tanya Q. Shang, Stuart E. Strand, Milton P. Gordon, Allison L. Moore, Sharon L. Doty, Suchitra Banerjee |
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| Rok vydání: | 2002 |
| Předmět: |
Agrobacterium
Gene Expression Bioengineering Plant Roots Applied Microbiology and Biotechnology Tissue culture Organ Culture Techniques Transformation Genetic Atropa belladonna Gene expression Botany Sense (molecular biology) Animals Transgenes biology Gene Transfer Techniques Cytochrome P-450 CYP2E1 biology.organism_classification Trichloroethylene Transformation (genetics) Biochemistry Hairy root culture Environmental Pollutants Rabbits Solanaceae Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 77:462-466 |
| ISSN: | 1097-0290 0006-3592 |
| Popis: | P450 2E1 is an important mammalian liver enzyme known to metabolize a wide range of compounds including several common environmental pollutants. The medicinal plant, Atropa belladonna, was transformed with Agrobacterium rhizogenes containing a binary vector with rabbit P450 2E1 in either the sense or antisense orientation. The resulting "hairy roots" were isolated and grown in liquid medium. Production of P450 2E1 protein was verified in the roots containing the 2E1 gene in the sense orientation. Transgenic and control root cultures were dosed with the environmental pollutant, trichloroethylene (TCE), and were analyzed for the TCE metabolites, chloral and trichloroethanol. The root cultures expressing the mammalian P450 2E1 had increased levels of the metabolites compared to the levels in the control roots. This method represents a quick way to screen transformants for expression of foreign genes before regeneration of whole plants, and also as a possible source of foreign protein for purification. |
| Databáze: | OpenAIRE |
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