Hurt D J, Winestock K D, O'Connor M L, Johnston M I
Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, MD 20814-4799.
Nucleic Acids Res. 1987 Nov 11;15(21):9057-73. doi: 10.1093/nar/15.21.9057.
Antisera elicited by immunization of rabbits with 4'-aminomethyl-trioxsalen (AMT)-modified poly(A,U) complexed with methylated bovine serum albumin was characterized in competition radioimmunoassays (RIA) and enzyme-linked immunosorbent assays (ELISA). AMT-poly(A,U) was over 10,000-fold more reactive than unmodified 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's in vitro and in intact cells.
用4'-氨甲基三氧杂蒽（AMT）修饰的聚（A，U）与甲基化牛血清白蛋白复合免疫兔子所产生的抗血清，在竞争放射免疫分析（RIA）和酶联免疫吸附分析（ELISA）中得到了表征。AMT-聚（A，U）的反应性比未修饰的聚（A，U）或单独的AMT高10000倍以上。该抗血清与AMT修饰的RNA和DNA有不同程度的交叉反应。AMT-尿苷的存在通常保证了强反应性。AMT的氨基对抗体结合有一定程度的贡献。高离子强度对结合没有显著影响，这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完整细胞中的细胞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具有可检测到的碱基对片段。这里描述的抗体将有助于研究体外和完整细胞中RNA的二级和三级结构。
