Molecular cloning and characterization of embryo-specific genes from 10-day embryos of rice (oryza sativa L. cv. tainung 67)zeng

• Main Authors: Xu, Hong-Ru, 許鴻儒
• Other Authors: Zeng, Meng-Jiao
• Format: Others
• Language: zh-TW
• Published: 1994
• Online Access: http://ndltd.ncl.edu.tw/handle/36372135316486605578

碩士 === 國立中興大學 === 分子生物研究所 === 82 === The objectives of the current research are to isolate the embryo- specific genes from 10-day embryos of rice, to examine the spatial- specific and temporal-specific expression of selected genes, and to study the genomic organization of these genes. A cDNA library was constructed from 10-day embryos of rice (Oryza sativa L. cv. Tainung 67). Eight-hundred and twenty-eight plaques that hybridized with [32-P]-labeled single- strand cDNA of 10-day embryos but not of mature embryos (45 days after pollination), were preliminary selected. Following in vivo excision of selected phages and repeating DNA dot blot hybridiation, 29 cDNA clones were further selected. The results of northern slot hybridization indicated that the the expression of 29 cDNA clones were higher in 10-day embryos than in 15-day embryos, and there little expression in the mature embryos or leaves of rice. The results of northern blot hybridization showed that the expression of pRE4、 pRE22、 pRE 、 pRE20 and pRE2 were highest in 9-day embryos among 7-, 10-, 11, and 15-day embryos, and there were little expressions in the mature embryos or leaves of rice. We were not able to detect any expression of selected genes in the 3-day seedling of rice, corn, azuki bean, pai-tsai, cauliflower, water convolvulus and potato. Thus, the data indicated that selected clones are 10-day embryos abundant and/or specific genes. According to 3''end nucleotide sequencing, the selected 29 cDNA clones could be divided into seven different genes or gene families. The results of Southern slot blot hybridization of rice genomic DNA indicated that the selected genes are the single or a few copies in the chromosome.