Cloning and Characterization of Rice Genomic DNAs of RE1M and RE31D

• Main Authors: Chien-Wei Hou, 侯建維
• Other Authors: Menq-Jiau Tseng
• Format: Others
• Language: zh-TW
• Published: 1999
• Online Access: http://ndltd.ncl.edu.tw/handle/84603671353409478077

碩士 === 國立中興大學 === 分子生物學研究所 === 87 === The objectives of the current research are to clone the genomic clones of RE1M and RE31D from 10-day embryos of rice, to analyze these two genes, and to study the regulation of RE31D gene expression. The genomic clones of RE1M and RE31DG were isolated from genomic library of 5-day etiolated shoot of rice selected by plaque hybridization with the cDNA probe of RE1M and RE31D, and named RE1MG and RE31DG, respectively. The RE1MG is 1,853 bp long, and contains a 1,590-bp ORF, a 262-bp 5' untranslated region, and a 334-bp 3' untranslated region, but no intron. The molecular weight of deduced amino acid of RE1MG is 43.7 kDa with 401 amino acids and a pI of 11.34. The identity of RE1MG is not known at this moment. The RE31DG is 3,877 bp long, and contains a 1,775-bp ORF, a 1666-bp 5' untranslated region, a 138-bp 3' untranslated region, and two introns. The molecular weight of deduced amino acid of RE31DG is 60.5 kDa with 541 amino acids and a pI of 11.99. Either the nucleotide sequence or the deduced amino acid sequence of RE31DG is highly homologous with the plant globulin gene. Two chimeric constructs contained 5' promoter region of RE31DG (-367~47 and -890~47) and GUS coding region were transferred to 10-day embryos, mature embryos and calli of rice via gene gun mediated transformation. The results of GUS histochemical staining after 2 days of transformation show that the transient expression of GUS in both constructs were high in 10-day and mature embryos, but low in the calli. The promoter of RE31DG has embryo-specific characters.