Two cDNA libraries were constructed from microdissected 214 rice proembryos (2-3 d after pollination) and 121 just differentiating young embryos (3-5 d after pollination) respectively through RT_PCR technique. The primary libraries had a total of 3.7×10 6 phages for the proembryos and a total of 2.5×10 6 phages for the just differentiating young embryos, in which 96% of the phages were recombinants. Insert sizes ranging from 400 bp to 3?500 bp were obtained. All of the above mentioned accorded with the general requirements of cDNA library construction.
为了监测钙调素 (Ca M)在植物细胞内的分布并探索其生物学功能 ,将水稻 Ca M和 GU S融合基因 (cam.gus)分别置于 35 S启动子和花粉特异启动子 L AT5 2 - 7控制下 ,构建出载体 p MD/ Ca M.GU S和 p BI/ L AT.Ca M.GU S,转化烟草 (N icotiana tobacum cv.SR ) ,GU S染色结果显示 ,Ca M.GU S融合蛋白广泛分布于植物根、茎、叶等组织 ,根尖生长点细胞、根毛细胞、表皮毛细胞、气孔保卫细胞、维管束细胞的细胞质中融合蛋白含量较多。花粉粒中也分布着 Ca M,并在萌发沟处表现出较高的浓度。保卫细胞和表皮毛细胞中 Ca M分布在相邻细胞连接处细胞模内侧 ,呈现出极性分布。研究结果表明 ,细胞内 Ca M的不均匀分布可能与细胞功能相关。