Schutz G, Kieval S, Groner B, Sippel A E, Kurtz D, Feigelson P
Nucleic Acids Res. 1977 Jan;4(1):71-84. doi: 10.1093/nar/4.1.71.
A procedure is presented for the purification of specific mRNAs, which exploits the ability of antibodies prepared against a native protein to bind to the nascent polypeptide on the polysome. Rather than precipitating these soluble antibody-polysome complexes with anti-antibody, which can lead to nonspecific trapping of polysomes, we have linked the anti-antibody to an insoluble matrix. Thus, the antibody-polysome complex binds to the anti-antibody support and nonspecific polysomes can easily be removed by several washes. We have found para-aminobenzyl cellulose (PAB cellulose), to be a suitable matrix for this purpose. This support can bind large quantities of anti-antibody and it displayed no detectable nonspecific affinity for polysomes or RNA. Using this procedure, we have obtained an apparently homogeneous preparation of ovalbumin mRNA.
本文介绍了一种纯化特定mRNA的方法，该方法利用针对天然蛋白质制备的抗体与多聚核糖体上新生多肽结合的能力。我们没有用抗抗体沉淀这些可溶性抗体-多聚核糖体复合物（这可能导致多聚核糖体的非特异性捕获），而是将抗抗体连接到不溶性基质上。因此，抗体-多聚核糖体复合物与抗抗体支持物结合，通过几次洗涤就可以很容易地去除非特异性多聚核糖体。我们发现对氨基苄基纤维素（PAB纤维素）是用于此目的的合适基质。这种支持物可以结合大量抗抗体，并且对多聚核糖体或RNA没有可检测到的非特异性亲和力。使用该方法，我们获得了明显均一的卵清蛋白mRNA制剂。
