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.
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