RECOMBINANT PINORESINOL/LARICIRESINOL REDUCTASE, RECOMBINANT DIRIGENT PROTEIN, AND METHODS OF USE

摘要

Dirigent proteins and pinoresinol/lariciresinol reductases have been isolated from Forsythia intermedia, Thuja plicata and Tsuga heterophylla, together with cDNAs encoding dirigent proteins and pinoresinol/lariciresinol reductases from these species. Accordingly, isolated DNA sequences are provided which code for the expression of dirigent proteins and pinoresinol/lariciresinol reductases. In other aspects, replicable recombinant cloning vehicles are provided which code for dirigent proteins or pinoresinol/lariciresinol reductases or for a base sequence sufficiently complementary to at least a portion of dirigent protein or pinoresinol/lariciresinol reductase DNA or RNA to enable hybridization therewith (e.g., antisense dirigent protein or pinoresinol/lariciresinol reductase RNA or fragments of complementary dirigent protein or pinoresinol/lariciresinol reductase DNA which are useful as polymerase chain reaction primers or as probes for genes encoding dirigent proteins or pinoresinol/lariciresinol reductases or related genes). In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding dirigent protein or pinoresinol/lariciresinol reductase. Thus, systems and methods are provided for the recombinant expression of dirigent proteins and/or pinoresinol/lariciresinol reductases that may be used to facilitate the production, isolation and purification of significant quantities of recombinant dirigent proteins and/or pinoresinol/lariciresinol reductases for subsequent use, to obtain expression or enhanced expression of dirigent proteins and/or pinoresinol/lariciresinol reductases in plants in order to enhance, or otherwise alter, lignan biosynthesis, or may be otherwise employed for the regulation or expression of dirigent proteins and pinoresinol/lariciresinol reductases.