一种与聚腺苷酸（poly(A)）反应的单克隆抗体的制备及其在聚腺苷酸阳性（poly(A)+）RNA 分离中的应用。

Production of a monoclonal antibody reactive with poly(A) and its application in the isolation of poly(A)+ RNA.

作者信息

Kitagawa Y, Ishibashi K

机构信息

Biotechnology Institute, Akita Prefectural College of Agriculture, Japan.

出版信息

J Immunol Methods. 1993 Aug 26;164(1):5-11. doi: 10.1016/0022-1759(93)90269-d.

DOI:10.1016/0022-1759(93)90269-d

PMID:7689620

Abstract

Hybridoma cells producing antibody reactive with poly(A) were isolated following fusion of spleen cells from a BALB/c mouse immunized with poly(A).poly(dT) and myeloma cells. The antibody was purified from ascitic fluid by the formation of immune complexes with poly(A). The antibody reacted with poly(A) and some homopolymers (poly(dT), poly(I) and poly(dC)) which was most probably due to its reaction with the ribose-phosphate part of the molecules, but not with cellular RNAs and DNA. Poly(A) tailed 5 S rRNA (about 20 nucleotides) and tobacco mosaic virus RNA (1060 nucleotides) were precipitated quantitatively by the antibody. Dot hybridization and in vitro translation confirmed that the RNA precipitated by the antibody from mouse spleen total RNA contained biologically active mRNA.

摘要

在用聚（A）·聚（dT）免疫的BALB/c小鼠的脾细胞与骨髓瘤细胞融合后，分离出了产生与聚（A）反应性抗体的杂交瘤细胞。通过与聚（A）形成免疫复合物，从腹水液中纯化出该抗体。该抗体与聚（A）和一些同聚物（聚（dT）、聚（I）和聚（dC））发生反应，这很可能是由于其与分子的核糖 - 磷酸部分发生反应，但不与细胞RNA和DNA反应。聚（A）尾的5S rRNA（约20个核苷酸）和烟草花叶病毒RNA（1060个核苷酸）可被该抗体定量沉淀。点杂交和体外翻译证实，抗体从小鼠脾脏总RNA中沉淀出的RNA含有生物活性mRNA。