Isolation of cytochrome P-450 cDNA clones from the higher plantCatharanthus roseusby a PCR strategy

✍ Scribed by Annemarie H. Meijer; Erik Souer; Robert Verpoorte; J. Harry C. Hoge

• Book ID: 104613775
• Publisher: Springer
• Year: 1993
• Tongue: English
• Weight: 478 KB
• Volume: 22
• Category: Article
• ISSN: 0167-4412
• DOI: 10.1007/bf00014944

✦ Synopsis

Cytochrome P-450 monooxygenases are membrane-bound enzymes involved in a wide range of biosynthetic pathways in plants. An efficient PCR strategy for isolating cytochrome P-450 cDNA clones from plant cDNA libraries is described. A set of degenerate primers for PCR amplification was designed to recognize nucleotide sequences specifying the highly conserved haembinding region of cytochrome P-450 proteins. Using this primer set and a non-specific primer, complementary to either the poly(A) tail of the cDNA clones or a phage vector sequence, we isolated 16 different cytochrome P-450 cDNA sequences from a cDNA library of Catharanthus roseus.