Isolation of specific messenger RNA by adsorption of polysomes to matrix-bound antibody.
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.
D000818:Animals; D000906:Antibodies; D002482:Cellulose / Q000031:analogs & derivatives; D055598:Chemical Phenomena; D002621:Chemistry; D002645:Chickens; D004372:Ducks; D005260:Female; D005914:Globins; D007136:Immunoglobulins; D009693:Nucleic Acid Hybridization; D010047:Ovalbumin / Q000096:biosynthesis*; D010057:Oviducts / Q000166:cytology*; D010058:Oviposition; D011061:Poly A / Q000032:analysis; D011132:Polyribosomes / Q000378:metabolism; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000302:isolation & purification*; D013045:Species Specificity
D000906:Antibodies; D007136:Immunoglobulins; D012333:RNA, Messenger; D011061:Poly A; D005914:Globins; D002482:Cellulose; D010047:Ovalbumin