Isolation of specific messenger RNA by adsorption of polysomes to matrix-bound antibody
نویسندگان
چکیده
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.
منابع مشابه
Evidence for nontranslated messenger fibonucleic acid in membrane-bound and free polysomes of rabbit liver.
In previous publications (SHAFRITZ, D. A., DRYSDALE, J. W., AND ISSELBACHER, K. J. (1973) J. Biol. Chem. 248, 3220-3227 and SHAFRITZ, D. A. (1974) J. Biol. Chem. 249, 81-88) we reported the isolation and translation of RNA fractions with messenger activity from membrane-bound and free polysomes of rabbit liver with evidence for specilic synthesis of albumin and ferritin in a reticulocyte cell-f...
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عنوان ژورنال:
- Nucleic acids research
دوره 4 1 شماره
صفحات -
تاریخ انتشار 1977