Mass isolation of pea nuclei.
Mass isolation of pea nuclei.
复制标题
豌豆核的质量分离。
DOI：
10.1104/pp.47.4.499
复制
发表时间：
1971
期刊：
影响因子：
7.4
通讯作者：
K. Tautvydas
中科院分区：
文献类型：
--
作者：
K. Tautvydas
传统过滤和离心程序的改进使得完整无细胞质豌豆 (Pisum sativum) 细胞核的产量从通常的 3% 提高到 10% 到 32%（每 7 克豌豆尖鲜重 6 x 10(8) 个细胞核），并在 80 到 90 分钟内完成分离。电子显微照片显示，分离的细胞核似乎保留了其结构完整性，并且在 20 摄氏度下至少 5 小时保持完整。分离的豌豆细胞核的 DNA:RNA:蛋白质比率为 3.1:1:9.9。通过将 (14)C 掺入来自 (14)C-UTP 的 RNA 来监测，它们的 RNA 聚合酶活性在约 10 分钟内呈线性，然后在接下来的 15 至 20 分钟内逐渐下降。
Improvements in conventional filtration and centrifugation procedures made it possible to increase the yield of intact cytoplasm-free pea (Pisum sativum) nuclei from the usual 3 to 10% to 32% (6 x 10(8) nuclei per 7 grams fresh weight of pea apices) and to complete the isolation in 80 to 90 minutes. The isolated nuclei appeared to retain their structural integrity as revealed in electron photomicrographs, and remained intact for at least 5 hours at 20 Celsius.The DNA:RNA: protein ratio of isolated pea nuclei was found to be 3.1:1:9.9. Their RNA polymerase activity, monitored by incorporation of (14)C into RNA from (14)C-UTP, was linear for about 10 minutes, and then gradually declined over the next 15 to 20 minutes.