Oakden K M, Azad A A, Lane B G
Can J Biochem. 1977 Jan;55(1):99-109. doi: 10.1139/o77-016.
(1) If wheat embryo 5S and 5.8S rRNA are differentially labelled, it can be shown that there is highly selective association of 5S [14C]RNA with 18S rRNA, and of 5.8S [3H]RNA with 26S rRNA when a solution (0.3 M NaCl) that contains approximately equimolar amounts of the four ribosomal polynucleotides is heated briefly (3 min) at 60 degrees C. (2) Comparison of Tm values and melting profiles for laboratory-prepared and natural 5.8S-26S rRNA hybrids suggests that restoration of the natural union between 5.8S and 26 rRNA can be achieved with facility and fidelity in the laboratory. (3) Union between 5.8S rRNA remains intact when wheat embryo ribosomes are disintegrated either by digestion with pronase or by treatment with sodium dodecyl sulphate, but the same treatments release 5S and 18S rRNA as freely migrating electrophoretic components. (4) Intact 18S and 26S rRNA can be prepared from small and large subunits, respectively, when wheat embryo ribosomes are dissociated by treatment with 0.5 M KCl. (5) Incidental to the principal investigation, it has been shown that, even after storage for more than 6 years at -70 degrees C, commercial supplies of roller-milled wheat germ yield S23 extracts that are very active in the cell-free translation of globin mRNA. (6) The physicochemical and possible biochemical significance of various types of intermolecular complexing between pairs of ribosomal polynucleotides is a subject of discussion.
(1) 如果对小麦胚5S和5.8S rRNA进行不同标记,结果表明,当含有大约等摩尔量的四种核糖体多核苷酸的溶液(0.3M NaCl)在60℃下短暂加热(3分钟)时,5S [14C]RNA与18S rRNA高度选择性结合,5.8S [3H]RNA与26S rRNA高度选择性结合。(2) 对实验室制备的和天然的5.8S - 26S rRNA杂交体的Tm值和熔解曲线进行比较,结果表明在实验室中可以方便且准确地实现5.8S和26S rRNA之间天然结合的恢复。(3) 当用链霉蛋白酶消化或用十二烷基硫酸钠处理使小麦胚核糖体解体时,5.8S rRNA之间的结合保持完整,但相同处理会释放出5S和18S rRNA作为自由迁移的电泳成分。(4) 当用0.5M KCl处理使小麦胚核糖体解离时,完整的18S和26S rRNA可以分别从小亚基和大亚基中制备出来。(5) 在主要研究过程中还发现,即使在-70℃下储存超过6年,市售的辊磨小麦胚芽制成的S23提取物在无细胞翻译珠蛋白mRNA方面仍具有很高的活性。(6) 核糖体多核苷酸对之间各种类型分子间复合的物理化学及可能的生化意义是一个讨论的主题。