从植物愈伤组织中提取多核糖体的要求。
Requirements for extraction of polyribosomes from plant callus cultures.
机构信息
Department of Botany and Plant Pathology, Michigan State University, East Lansing, Michigan 48824.
出版信息
Plant Physiol. 1977 May;59(5):800-2. doi: 10.1104/pp.59.5.800.
A procedure was developed to isolate polyribosomes from plant cell cultures. Tobacco callus extracted in 10 mm MgCl(2), 80 mm KCl, 250 mm sucrose, and 140 mm tris-HCl (pH 8.2) yielded larger amounts of polysomes than cells extracted in higher or lower ionic strength or pH buffers. Optimal conditions for extraction of polysomes from soybean callus were identical except the most suitable pH for recovery was 8.5. Addition of the divalent cation chelator, ethylene glycol-bis(2-aminoethyl ether)-tetraacetic acid (EGTA) to the extraction medium improved polysomal yield from tobacco and soybean cultures. Polysomes were successfully extracted from potato, tomato, corn, and barley cell cultures in extraction medium supplemented with EGTA.
我们开发了一种从植物细胞培养物中分离多核糖体的方法。在 10 mM MgCl(2)、80 mM KCl、250 mM 蔗糖和 140 mM tris-HCl(pH 8.2)中提取的烟草愈伤组织比在更高或更低离子强度或 pH 缓冲液中提取的细胞产生更多的多核糖体。从大豆愈伤组织中提取多核糖体的最佳条件相同，只是最适合回收的 pH 值为 8.5。在提取培养基中添加二价阳离子螯合剂乙二醇双(2-氨基乙基醚)四乙酸(EGTA)可提高烟草和大豆培养物中多核糖体的产量。在添加 EGTA 的提取培养基中，成功地从马铃薯、番茄、玉米和大麦细胞培养物中提取了多核糖体。
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