Curtis P J, Mantei N, van den Berg J, Weissmann C
Proc Natl Acad Sci U S A. 1977 Aug;74(8):3184-8. doi: 10.1073/pnas.74.8.3184.
Dimethyl sulfoxide-induced Friend cells were labeled for periods of 5-60 min. The denatured RNA was fractionated by sucrose gradient centrifugation and the distribution of alpha- and beta-globin-specific [(3)H]RNA was determined by hybridization to hybrid plasmids containing mouse alpha- and beta-globin DNA, respectively. After 5 min of labeling, a 15S peak of beta-globin-specific (but not alpha-globin-specific) [(3)H]RNA was detected, next to an equal amount of 10S beta-globin [(3)H]RNA. With increasing periods of labeling, the amount of 15S beta-globin [(3)H]RNA remained constant but the amount 10S beta-globin [(3)H]RNA increased steadily. alpha-Globin-specific [(3)H]RNA sedimented at 11 S after 5 min of labeling and at 9.5 S after longer labeling periods. Analysis of 15S globin-specific [(3)H]RNA purified by the poly(dC)-cDNA method [Curtis, P. J. & Weissmann, C. (1976) J. Mol. Biol. 106, 1061-1075] showed oligonucleotides characteristic of beta-globin mRNA but not of alpha-globin mRNA, as well as about 20 new oligonucleotides. Our results suggest that 10S beta-globin mRNA arises via a 15S precursor that has a half-life of 5 min or less; 9.5S alpha-globin mRNA may be derived from an 11S precursor.
用二甲基亚砜诱导的Friend细胞标记5 - 60分钟。将变性RNA通过蔗糖梯度离心进行分级分离,并分别通过与含有小鼠α - 和β - 珠蛋白DNA的杂交质粒杂交来确定α - 和β - 珠蛋白特异性[³H]RNA的分布。标记5分钟后,在等量的10Sβ - 珠蛋白[³H]RNA旁边检测到一个15S峰的β - 珠蛋白特异性(而非α - 珠蛋白特异性)[³H]RNA。随着标记时间的增加,15Sβ - 珠蛋白[³H]RNA的量保持恒定,但10Sβ - 珠蛋白[³H]RNA的量稳步增加。α - 珠蛋白特异性[³H]RNA在标记5分钟后沉降在11S,标记时间更长后沉降在9.5S。通过聚(dC) - cDNA方法[柯蒂斯,P. J. & 魏斯曼,C.(1976年)《分子生物学杂志》106,10,61 - 1075]纯化的15S珠蛋白特异性[³H]RNA分析显示,其寡核苷酸具有β - 珠蛋白mRNA而非α - 珠蛋白mRNA的特征,以及约20种新的寡核苷酸。我们的结果表明,10Sβ - 珠蛋白mRNA通过半衰期为5分钟或更短的15S前体产生;9.5Sα - 珠蛋白mRNA可能来自11S前体。