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通过 Gus 基因 在谷子 胚性愈伤 组织中 的瞬时表 达检测, 分析 了影 响谷 子愈 伤组 织基 因枪转化效 率的几个 因素。钨 粉的包 被质粒 D N A 用量以 3 μg/ m g 钨粉 , Ca Cl2 浓度15 m ol/ L,亚 精胺浓度40 m m ol/ L 为最 佳,过高 的 D N A 、 Ca Cl2 和亚精 胺加入量 均显著降 低转 化率 ; J Q700 基因 枪样品室高度 7 c m ,轰 击速 度 400 ~450 m /s 转 化, 较其 它 速度 和样 品 室高 度下 转 化 Gus 基因 表 达率高;每 次转化的 愈伤组织 用量对 转化效率 也有影 响, 以1 ~2 g Gus 基 因表 达斑 最多; 玉米 ubiquitin启动子 较 Ca M V35 S 启动子 的瞬时表 达显著增 强。
Through the transient expression of Gus gene in embryogenic callus of millet, several factors influencing the efficiency of gene gun transformation in millet callus were analyzed. The dosage of coating powder D N A for tungsten powder was the best with 3 μg / m 3 tungsten powder, CaCl 2 concentration 15 m ol / L and spermidine concentration 40 m mol / L, and excessive D N A , CaCl 2 and spermidine significantly reduced the conversion rate; J Q700 gene gun sample chamber height 7 c m bombardment speed 400 ~ 450 m / s transformation, compared with other rates and the height of the sample chamber transformed Gus gene expression The transformation efficiency was also affected by the amount of callus transfered in each transformation, with 1 ~ 2 g Gus gene expression spot the most; the transient expression of ubiquitin promoter in maize was significantly higher than that of CaM V35 S promoter.