Plant, Animal & Microbe Genomes X Conference

January 12-16, 2002
Town & Country Convention Center
San Diego, CA

Poster: Genome Sequencing & ESTs
            

MOLECULAR CLONING OF RICE ENDOSPERM SPECIFIC GENES AT CELL DIFFERENTIATION STAGE FROM THE SUBTRACTIVE cDNA LIBRARY

¹ Laboratory of Genetic Engineering, Faculty of Agriculture, Kyoto Prefectural University, Nakaragi-cho, Shimogamo, Sakyo, Kyoto, 606-8522, Japan

² Laboratory of Biophysical Chemistry, Graduate School of Aagriculture and Biological Science, Osaka Prefectural University, Gakuen-cho, Sakai, Osaka, 599-8531, Japan

Rice aleurone layer and starchy endosperm tissue are originated in a endosperm mother cell. Aleurone layer is very different from starchy endosperm tissue structurally and physiologically. We are interested in cell differentiation mechanisms of endosperm tissue. Then we would like to clone the functional genes from the differentiation endosperm cell. The cell differentiation of endosperm is started at 3 days after flowering (3DAF) and ended at 7DAF. Then we synthesized cDNA from endosperm tissues at 3DAF, and the cDNAs at 7DAF were subtracted from the former cDNAs. So, we constructed the subtraction cDNA library at 3DAF. We screened the cDNA library by using the single strand cDNAs at 3DAF as a probe. We picked up about two hundred positive clones and analyzed the positive clone sequences, so we got the partial cDNA sequences (including 3'UTR sequence) from the candidate clones. These cDNA sequences were analyzed using BLAST program, and twenty percent of the candidate clones corresponded to registered rice genome sequences. And a half of these sequences corresponded to the predicted gene sequences using the virtual transcription sequence database (VTS-database ver.8) , which was estimated from rice genome sequence data using GENESCAN program. We were successful in the cloning of some full-length cDNAs, which were expressed in endosperm tissue at cell differentiation stage.