Rzepecki R, Markiewicz E, Szopa J
Institute of Biochemistry, Wrocław University, Poland.
Acta Biochim Pol. 1995;42(2):171-6.
The nuclear matrices from White bush (Cucurbita pepo var. patisonina) cell nuclei have been isolated using three methods: I, standard procedure involving extraction of cell nuclei with 2 M NaCl and 1% Triton X-100; II, the same with pre-treatment of cell nuclei with 0.5 mM CuSO4 (stabilisation step); and III, method with extraction by lithium diiodosalicylate (LIS), and compared the polypeptide pattern. The isolated matrices specifically bind SAR DNA derived from human beta-interferon gene in the exogenous SAR binding assay and in the gel mobility shift assay. Using IgG against the 32 kDa endonuclease we have found in the DNA-protein blot assay that this protein is one of the proteins binding SAR DNA. We have identified three proteins with molecular mass of 65 kDa, 60 kDa and 32 kDa which are responsible for SAR DNA binding in the gel mobility shift assay experiments.
已使用三种方法从白布什南瓜（西葫芦变种帕蒂索尼纳）细胞核中分离出核基质：方法一，采用标准程序，即用2M氯化钠和1% Triton X-100提取细胞核；方法二，同样用0.5 mM硫酸铜对细胞核进行预处理（稳定化步骤）；方法三，用二碘水杨酸锂（LIS）提取的方法，并比较了多肽图谱。在外源SAR结合试验和凝胶迁移率变动试验中，分离出的核基质能特异性结合源自人β-干扰素基因的SAR DNA。在DNA-蛋白质印迹试验中，使用针对32 kDa核酸内切酶的IgG，我们发现该蛋白是结合SAR DNA的蛋白之一。在凝胶迁移率变动试验中，我们鉴定出三种分子量分别为65 kDa、60 kDa和32 kDa的蛋白，它们负责结合SAR DNA。
