含聚腺苷酸的多瘤“早期”和“晚期”信使核糖核酸的分离与特性分析
Isolation and characterization of poly(A)-containing polyoma "early" and "late" messenger RNAs.
作者信息
Rosenthal L J
出版信息
Nucleic Acids Res. 1976 Mar;3(3):661-76. doi: 10.1093/nar/3.3.661.
Abstract
During late lytic infection of mouse kidney cell cultures polyoma 16S and 19S (late 19S RNA) were isolated by oligo(dT)-cellulose chromatography. Approximately 60-80% of total cytoplasmic polyoma RNA contained tracts of poly(A) which were retained by oligo(dT)-cellulose. Early in lytic infection when viral DNA synthesis and the production of capsid protein are blocked by the addition of 5-fluorodeoxyuridine, approximately 100% of polyoma "early" 19S RNA was quantitatively retained by oligo(dT)-cellulose indicating the presence of poly(A) tracts on most 19S mRNA molecules. In addition, 2 classes polyoma RNA, synthesized after the onset of cellular RNA synthesis under conditions where DNA synthesis is inhibited with 5-fluorodeoxyuridine, were found to contain tracts of poly(A). These species sedimenting at 16S and 19S in aqueous sucrose density gradients were also quantitatively retained by oligo (dT)-cellulose.
摘要
在小鼠肾细胞培养物的晚期裂解感染期间，通过寡聚（dT）-纤维素色谱法分离出多瘤病毒16S和19S（晚期19S RNA）。大约60-80%的细胞质多瘤病毒RNA含有多聚（A）序列，这些序列被寡聚（dT）-纤维素保留。在裂解感染早期，当通过添加5-氟脱氧尿苷阻断病毒DNA合成和衣壳蛋白产生时，大约100%的多瘤病毒“早期”19S RNA被寡聚（dT）-纤维素定量保留，这表明大多数19S mRNA分子上存在多聚（A）序列。此外，在DNA合成被5-氟脱氧尿苷抑制的条件下，在细胞RNA合成开始后合成的两类多瘤病毒RNA被发现含有多聚（A）序列。这些在蔗糖密度梯度水溶液中沉降系数为16S和19S的RNA种类也被寡聚（dT）-纤维素定量保留。
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Purification of polyoma virus.多瘤病毒的纯化
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