Isolation and in vitro assembly of nuclear ribonucleoprotein particles and purification of core particle proteins.

摘要: Publisher Summary This chapter describes the isolation and in vitro assembly of nuclear ribonucleoprotein particles purification core particle proteins. In order to obtain hnRNP complexes which are not contaminated with ribosomes or other cytoplasmic components it is very important rapidly nuclei free material. Native C-protein tetramers can be purified from isolated 40 S through a two-step procedure using gel filtration 350 mM NaCI anion-exchange chromatography. Many questions regarding composition, structure, function have been difficult answer experimentally because RNA component monoparticles as heterogeneous entire complement pre-mRNA proteins do protect packaged length endogenous nuclease action. Although will dissociate 0.7 M NaC1 spontaneously reassemble on back-dialysis into low salt, this complicated by necessity removing fragments under high-salt conditions prevent reassembly.