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肌动蛋白位于HeLa细胞的细胞核和核基质中。

[Actin is located in the nucleus and nuclear matrix of HeLa cells].

作者信息

Wang H, Xing M

机构信息

Molecular Biology Department, Bethune University of Medical Sciences, Changchun 130021, China.

出版信息

Yi Chuan Xue Bao. 2001;28(3):211-6.

Abstract

HeLa cell nuclei were isolated and the nuclear matrix specimens were prepared. After labelled with an anti-actin antibody and FITC-conjugated secondary antibody, both the nuclei and nuclear matrix specimens were observed to emanate specific yellow-green fluorescence. The fluorescent signals in the nuclear matrix were much stronger than that in the nuclei, and the signals in former were widespread throughout the whole structure while that in the latter were mainly defined to their peripheral regions. A 43 kD band was revealed in nuclei and nuclear matrix specimens by SDS-PAGE, and was then proved to be actin by Western blot, confirming the immuno-fluorescence observations. When stained with TRICT-conjugated phalloidin, both the nuclei and nuclear matrix specimens were found to give off specific, red fluorescent signals which represent the location of F-actin (filamentous actin), and the pattern of TRITC signals distribution in the nuclei and nuclear matrix showed similarity with that of FITC signals. The existence and significance of actin and F-actin in the nuclei and nuclear matrix were discussed.

摘要

分离出HeLa细胞核并制备核基质标本。用抗肌动蛋白抗体和异硫氰酸荧光素(FITC)偶联的二抗标记后,观察到细胞核和核基质标本均发出特定的黄绿色荧光。核基质中的荧光信号比细胞核中的强得多,前者的信号遍布整个结构,而后者的信号主要集中在其周边区域。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)在细胞核和核基质标本中显示出一条43 kD的条带,随后通过蛋白质免疫印迹法证实其为肌动蛋白,这与免疫荧光观察结果一致。用四甲基罗丹明异硫氰酸酯(TRITC)偶联的鬼笔环肽染色时,发现细胞核和核基质标本均发出代表丝状肌动蛋白(F-肌动蛋白)位置的特定红色荧光信号,并且TRITC信号在细胞核和核基质中的分布模式与FITC信号相似。讨论了肌动蛋白和F-肌动蛋白在细胞核和核基质中的存在及其意义。

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