摘要
自烟草悬浮培养细胞中分离出细胞核,提取核总蛋白后,用DNase I消化去除核酸和2MNaCl抽提后制备成细胞核骨架.以抗β-actin的抗体作一抗,辣根过氧化物酶标记(HRP)的羊抗兔抗体作二抗进行免疫印迹.结果证实,在细胞质、细胞核与核骨架中均存在与肌动蛋白抗体呈阳性反应的抗原.以抗myosinⅠβ抗体作一抗,辣根过氧化物酶(HRP)标记的羊抗兔抗体作二抗进行免疫印迹.也证实了细胞核中存在与肌球蛋白抗体呈阳性反应的抗原.上述结果证明,肌动蛋白和肌球蛋白是烟草悬浮细胞细胞核及核骨架的组成成分.来源于老鼠纤维原细胞细胞核中的肌球蛋白I存在于烟草细胞核中也表明细胞核中的肌动蛋白和肌球蛋白在生物进化过程中是相当保守的.
Isolation the nuclei from the suspension cells of tobacco （ Nicotiana tabacum L. ）, then the total nuclear proteins were extracted by nuclear-cytosol extraction kit. Nuclear matrix was obtained by digesting and extracting, respectively with DNase Ⅰ and 2M NaC1. The results of the immunoblot of anti-β-actin antibody as immunomarker indicated that the cytoplasm, nuclei andnuclear matrix had positive reaction with the antibody. Similarly, immunoblot tests also showed that the nuclei reacted positively with the anti-NM Ⅰ β antibody. The result of the two immunoblot experiments verified the presence of β-actin was the constituent of the nuclei and nuclear matrix. Simultaneously, nuclear myosin Ⅰ β （NM Ⅰ β） was also a constituent of the nuclear proteins. It displayed that the NM Ⅰ β which originated from the nuclei of the mouse fibroblasts located in the nuclei of the suspension cells of tobacco. Furthermore, it demonstrated that the conservation of the β-actin and NM I β during the biology evolution.
出处
《首都师范大学学报（自然科学版）》
2008年第1期55-60,74,共7页
Journal of Capital Normal University：Natural Science Edition
关键词
烟草悬浮细胞
细胞核
核骨架
肌动蛋白
肌球蛋白
suspension cell of tobacco （ Nicotiana.tabacum L. ）, nucleus, nuclear matrix, actin, myosin.
