Rivera Maria C, Maguire Bruce, Lake James A
Department of Biology and Center for the Study of Biological Complexity, Virginia Commonwealth University, Richmond, Virginia 23284;
Primary Pharmacology Group, Pfizer Global Research and Development, Groton, Connecticut 06340;
Cold Spring Harb Protoc. 2015 Apr 1;2015(4):363-7. doi: 10.1101/pdb.prot081372.
Structural and functional studies of ribosomal subunits require the dissociation of intact ribosomes into individual small and large ribosomal subunits. The dissociation of the prokaryotic 70S ribosomes into the 50S and 30S subunits is achieved by dialysis against a buffer containing a lower Mg(2+) concentration. Eukaryotic 80S ribosomes are dissociated into 60S and 40S subunits by incubation in a buffer containing puromycin and higher KCl and Mg(2+) concentrations.
核糖体亚基的结构和功能研究需要将完整的核糖体解离成单个的小核糖体亚基和大核糖体亚基。通过用含有较低镁离子浓度的缓冲液进行透析，可将原核生物的70S核糖体解离成50S和30S亚基。真核生物的80S核糖体通过在含有嘌呤霉素以及较高氯化钾和镁离子浓度的缓冲液中孵育而解离成60S和40S亚基。
