Fernando M A, Pasternak J
Parasitology. 1977 Feb;74(1):27-32. doi: 10.1017/s0031182000047508.
A procedure is described for the preparation of purified nuclei of chick intestinal cells infected with Eimeria necatrix. During second-generation schizogony the host-cell membrane becomes resistant to homogenization, hypotonic shock and ultrasonication. However, brief treatment with a low concentration of trypsin (0.001%) will disrupt the cell membrane. Thereafter the nuclei can be selectively harvested from contaminating nuclear types and schizonts by a series of centrifugations. Electron microscopy and Feulgen-DNA microspectrophotometry showed that the nuclear preparations retained their integrity with little, if any, loss of DNA. Chemical determinations confirmed that host-cell nuclear DNA synthesis was stimulated by coccidial infection and further established that quantitatively there was a coincident increase in the amount of host nuclear RNA during schizogony. It is now feasible to examine directly the effects of coccidial infection upon the host's nuclear apparatus.
本文描述了一种制备感染柔嫩艾美耳球虫的鸡肠道细胞纯化细胞核的方法。在第二代裂殖生殖过程中，宿主细胞膜对匀浆、低渗休克和超声处理具有抗性。然而，用低浓度胰蛋白酶（0.001%）进行短暂处理会破坏细胞膜。此后，通过一系列离心可以从污染的核类型和裂殖体中选择性地收获细胞核。电子显微镜和福尔根 - DNA显微分光光度法表明，核制剂保持了其完整性，DNA几乎没有损失（如果有损失的话）。化学测定证实，球虫感染刺激了宿主细胞核DNA的合成，并进一步确定在裂殖生殖过程中宿主核RNA的量在数量上同时增加。现在直接研究球虫感染对宿主核装置的影响是可行的。
