Production of Useful Compounds by Genetic Transformation

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(1)The Genes of Flavonoids biosynthesisThe genes of flavonoids biosynthesis are induced by environmental stimuli. This induction was regulated at transcriptional level. In this project we focussed on the genes of chalcone synthase(CHS)and reductase(CHR)co-acting with chalcone synthase.The cDNA of CHS was cloned from Pueraria lobata cDNA library. And the cDNA of reductase was cloned by PCR. The genome genes of CHS and CHR were cloned from genome DNA library using the each cDNA as probe.(2)The Genes of Terpenoids BiosynthesisThe biosynthesis pathway of triterpene and sterol is dissected at cyclization of oxidosqualene in plant. Cycloartenol cyclase produces cycloartenol from oxidosqualene and leads to sterols. On the other hand, beta-amyrin cyclase, one of triterpene cyclases, cyclase oxidosqualene to beta-amyrin and leads to triterpines. In this project we focussed on the genes of cycloartenol cyclase and beta-amyrin cyclase.These two enzymes were purified from pea, and several amino acid sequences were determined. On the basis of these sequences, the oligo nucleotides were synthesized and used as primer of PCR. The partial length cDNA was obtained, and cDNA library was screened using the obtained partial length cDNA as probe. The full length cDNA of beta-amyrin cyclase was obtained, but cDNA of cycloartenol cyclase has not been obtained full length cDNA yet.(3)Introduction of Foreign Genes and Analysis of PromotersThe promoter region of beta-conglycinin gene which is coding the storage protein of soybean seeds is analyzed by construction of deletion clones. The sequence of "CATGCAT" is indispensable for specific expression in seeds.In the research of introduction of foreign genes into plant, some new results about effects of adjacent sequences, effects of transferred location and effects of double transformation, were obtained.