Highly purified micro- and macronuclei from Tetrahymena thermophila isolated by percoll gradients
Highly purified micro- and macronuclei from Tetrahymena thermophila isolated by percoll gradients
复制标题
通过 Percoll 梯度从嗜热四膜虫中分离出高度纯化的微核和大核
DOI：
--
复制
发表时间：
1983
期刊：
影响因子：
--
通讯作者：
M. Swancutt
中科院分区：
文献类型：
--
作者：
S. Allen;T. C. White;J. Langmore;M. Swancutt
A new procedure is described that utilizes Percoll gradients for purifying micronuclei (MIC) and macronuclei (MAC) from Tetrahymena therrnophila. Separation of MIC from MAC during certrifugation in Percoll gradients occurs as a result of their difference in size rather than density. Three kinds of tests were used to evaluate the purity of the nuclei: visualization of the nuclei by light microscopy; examination of the nuclei by electron microscopy; and Southern blots of MIC and MAC DNA probed with the 5s rRNA genes or a fragment from the MAC extrachromosomal rDNA molecule. When examined under the light microscope, the isolated MIC and MAC have much lower nuclear cross contamination levels than previous methods have reported. MIC's contaminated with less than 1 MAC in 1000 MIC and MAC'S contaminated with less than 1 MIC in 500 MAC can be routinely prepared. Quantitative analyses of electron micrographs gave higher estimates of cross contamination in our purified nuclei, which may, in part, be explained by the difficulty in identifying small MIC or MAC fragments. Southern blots of MIC and MAC DNA probed with 5s rDNA confirmed the level of MAC contamination in the MIC estimated by light microscopy during purification of the nuclei. The level of nucleolar contamination in the MIC was estimated at 10% by Southern blots of MIC and MAC DNA, derived from a heterokaryon with distinctive MIC and MAC Barn HI sites, using an rDNA probe. HE ciliate Tetrahymena thermophila has two kinds ofnuclei
DOI：
10.1073/pnas.78.12.7436
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
YAO, MC;YAO, CH
通讯作者：
YAO, CH