PAG-II Plant Genome II Conference

Town & Country Conference Center, San Diego, CA, January, 1994.


PG-II: EVALUATION OF cDNA LIBRARIES FOR SPECIFIC GENE CLONING

EVALUATION OF cDNA LIBRARIES FOR SPECIFIC GENE CLONING.

Hirofumi Uchimiya, Institute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo 113, Japan.


Recent developments in recombinant DNA technology have made it possible to isolate and characterize plant genes. Generally, such methods rely on the availability of either specific antibodies or appropriate oligonucleotides to target genes. We have carried out random nucleotide sequencing of cDNA clones. We have chosen the graminaceous monocot rice (Oryza sativa L.) as the experimental material, because classical genetics has mapped numerous morphological and isozyme markers to their respective chromosomes. To achieve the specific gene identification, our strategy has been based on the construction of cDNA libraries from cultured cells [Uchimiya, H. et al., Plant Journal, 2, 1005-1009 (1992)], which had been grown under stressed conditions such as salts, high osmotic pressures, and amino acid depletion. Partially resolved nucleotide sequences were used to search the GenBank database for nucleotide sequence homology. Here, we show that a number of stress-related genes can be identified from the cDNA libraries. Potential application of such cDNA clones for the study of molecular mechanism underlying stress responses of rice plants using genes encoding adenylate kinase [Kawai, M. et al., Plant Journal, 2, 845-854 (1992)], NDP kinase [Yano, A. et al., Plant Mol. Biol., in press (1993)], small GTP binding protein [Kidou, S. et al., FEBS Lett., 332, 282-286 (1993)], 14-3-3 protein [Kidou, S. et al., Plant Mol. Biol., 21, 191-194 (1993)] and other proteins will be reported.


Return to Previous Page or Intl-PAG Homepage