Antibody-nucleic acid interactions. Antibodies to psoralen-modified RNA as probes of RNA structure.
Antibody-nucleic acid interactions. Antibodies to psoralen-modified RNA as probes of RNA structure.
复制标题
抗体-核酸相互作用。
DOI：
10.1093/nar/15.21.9057
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发表时间：
1987
影响因子：
14.9
通讯作者：
Johnston,MI
中科院分区：
文献类型：
--
作者：
Hurt,DJ;Winestock,KD;O'Connor,ML;Johnston,MI
通过竞争性放射免疫测定（RIA）和酶联免疫吸附测定（ELISA）对用与甲基化牛血清白蛋白复合的4'-氨甲基三氧沙林（AMT）修饰的聚（A，U）进行免疫的兔子产生的抗血清进行了表征。 AMT-聚(A,U) 的反应性比未改性聚(A,U) 或单独的AMT 高10,000 倍以上。抗血清与 AMT 修饰的 RNA 和 DNA 发生不同程度的交叉反应。 AMT-尿苷的存在通常保证了强烈的反应性。 AMT 的氨基对抗体结合有较小程度的贡献。高离子强度对结合没有显着影响，表明结合不涉及离子对的形成。鼠脑心肌炎病毒复制中间体以及细胞RNA和DNA在完整细胞中被补骨脂素修饰，这表明完整细胞中的EMCV RNA和细胞RNA具有可检测的碱基对延伸。这里描述的抗体可用于研究体外和完整细胞中 RNA 的二级和三级结构。
Antisera eliclted by immunization of rabbits wlth 4'-aminomethyl-trioxsalen (AMT)-modified poly(A,U) complexed with methylated bovine serum albumin was characterized in competition radiolmmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). AMT-poly(A,U) was over 10,000-fold more reactive than unmodlfied poly(A,U) or AMT alone. The antiserum cross-reacted to varying extents with AMT-modified-RNA's and -DNA's. The presence of AMT-uridine usually assured strong reactivity. The amino group of AMT contributed to antibody binding to a small degree. Binding was not significantly affected by high ionic strength, suggesting that binding does not involve ion pair formation. Murine encephalomyocarditis virus replicative intermediates, as well as cellular RNA and DNA were modified by psoralen in intact cells, suggesting that EMCV RNA and cellular RNA's in intact cells possess detectable stretches of base pairs. The antibodies described here will be useful in studying the secondary and tertiary structure of RNA'sin vitroand in intact cells.