Kang Y J, Olson M O, Jones C, Busch H
Cancer Res. 1975 Jun;35(6):1470-5.
The nucleolar acid-soluble proteins from normal rat liver and Novikoff hepatoma ascites cells were labeled in vivo for 2 hr after the injection of [32P]orthophosphate and in vitro with [gamma-32P]adenosine triphosphate in systems containing 0.25 M sucrose, 5 mM MgCl2, 12.5 mM NaCl, and 0.05 M Tris-HCl buffer, pH 7.3, AT 37 DEGREES. Two-dimensional polyacrylamide gel electrophoresis and autoradiography showed that approximately 40 and 20 protein spots were labeled in vivo and in vitro, respectively. There were some marked differences in labeling in vivo between normal rat liver and Novikoff hepatoma acid-soluble nucleolar proteins. By 32P analysis of gel slices, the proportion of the total 32P incorporated into protein Spot A1-4 was greater in normal liver, and the proportion of 32P incorporated into some high-molecular-weight protein spots, such C23-24 and C26-27, was greater in the Novikoff hepatoma ascites cells. With the in vitro incubation system, the 32P uptake per mg protein was about twice as high in Novikoff hepatoma nucleolar proteins as in normal rat liver nucleolar proteins but, generally, the same proteins were labeled in both tissues.
在注射[32P]正磷酸盐后，对正常大鼠肝脏和诺维科夫肝癌腹水细胞中的核仁酸溶性蛋白质进行2小时的体内标记，并在含有0.25M蔗糖、5mM氯化镁、12.5mM氯化钠和0.05M Tris-HCl缓冲液（pH 7.3）的体系中，于37摄氏度用[γ-32P]三磷酸腺苷进行体外标记。二维聚丙烯酰胺凝胶电泳和放射自显影显示，体内和体外分别有大约40个和20个蛋白质斑点被标记。正常大鼠肝脏和诺维科夫肝癌酸溶性核仁蛋白质在体内标记方面存在一些显著差异。通过对凝胶切片进行32P分析，在正常肝脏中，掺入蛋白质斑点A1-4的总32P比例更高，而在诺维科夫肝癌腹水细胞中，掺入一些高分子量蛋白质斑点（如C23-24和C26-27）的32P比例更高。在体外孵育体系中，诺维科夫肝癌核仁蛋白质每毫克蛋白质的32P摄取量约为正常大鼠肝脏核仁蛋白质的两倍，但一般来说，两个组织中被标记的是相同的蛋白质。
