| 免疫亲和层析法分离纯化含次黄嘌呤核苷mRNA |
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| 引用本文： | 罗晓星,杨静华.免疫亲和层析法分离纯化含次黄嘌呤核苷mRNA[J].生物技术通讯,2002,13(3):179-182. |
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| 作者姓名： | 罗晓星 杨静华 |
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| 作者单位： | 1. 第四军医大学药理学教研室,西安,710032
2. Section of Trauma and Cntical Care,SurgeryDepartment Yale University School of Medicine,New Haven,CT 06520,USA |
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| 基金项目： | 美国耶鲁大学医学院OSHE基金 |
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| 摘 要： | 采用免疫亲和层析法分离纯化含次黄嘌呤核苷的mRNA(I-mRNA）。将Poly-I与BSA交联，并用Poly-I-BSA交联体免疫家兔，获取抗次黄呤核苷抗血清，斑点印迹法检测到该抗体滴度高、选择性强，将抗次黄嘌呤核苷抗体与蛋白A Sapharose交联，并制备抗体亲和层析柱。将小鼠肺mRNA上样于层析柱，淋洗层析柱后，用洗脱液洗脱I－mRNA，并以斑点印迹法在洗脱液中检测到很强的I－mRNA阳性信号；淋洗液中I－mRNA阳性信号的强度则随淋洗液体积的增加而减弱。以上结果表明，应用本方法可以有效分离得到含次黄嘌呤核苷的mRNA。
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| 关 键 词： | 免疫亲和层析法 分离纯化 RNA编辑 次黄嘌呤核苷 抗次黄嘌呤核苷抗体 |
| 文章编号： | 1009-0002(2002)03-0179-04 |
| 修稿时间： | 2002年1月23日 |
Immunoaffinity purification of inosine-containing mRNA on colulnn of anti-inosine antibody |
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| Reuven Rabinovici.Immunoaffinity purification of inosine-containing mRNA on colulnn of anti-inosine antibody[J].Letters in Biotechnology,2002,13(3):179-182. |
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| Authors: | Reuven Rabinovici |
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| Abstract: | Inosine containing mRNA was purified by immunoaffinity chromatograph on the anti-inosine antibody col-umn.Poly(I)was coupled to BSA and the conjugate was used for immunizing rabbit in order to obtain anti-inosine serum.The tide and specificity of anti-inosine antibody were determined by dot blot assay and the results showed that anti-inosine antibody at1:10,000dilution had strong signal at poly(I)spots but no any cross reaction with poly(A),had very weak cross reaction with high loading amount of poly(G)spot(1000ng).The strong signals of I-mR-NA were detected from the elution of anti-inosine antibody column while there was very weak signal in last fraction of wash buffer.The results indicated that inosine containing mRNA could be successfully isolated from mRNA by using immunoaffinity chromatograph. |
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| Keywords: | RNA editing inosine anti-inosine antibody immunoaffinity purification |
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