Van Etten J L, Vidaver A K, Koski R K, Semancik J S
J Virol. 1973 Sep;12(3):464-71. doi: 10.1128/JVI.12.3.464-471.1973.
The Pseudomonas phaseolicola bacteriophage phi6 incorporated labeled UTP into an acid-insoluble precipitate. Incorporation was dependent on the presence of manganese acetate, ATP, GTP, CTP, and a short heat treatment of the phage; the reaction was stimulated by NH(4)Cl. The substitution of (14)C-ATP, -CTP or -GTP for UTP, together with the appropriate unlabeled ribonucleoside triphosphates, disclosed that CMP was incorporated to the greatest extent followed by GMP, UMP, and AMP. Radioactive RNAs formed by the reaction were resistant to RNases A and T(1) in high salt but susceptible to these nucleases in low salt. The labeled RNA co-sedimented and co-electrophoresed with phi6 double-stranded (ds) RNA. However, the distribution of the radioactivity into the three ds-RNA components varied depending on the (14)C-ribonucleoside triphosphate used in the reaction. The incorporation of UMP was primarily into the two smaller ds-RNA segments, GMP primarily into the large ds-RNA segment, and CMP and AMP were about equally distributed into all three ds-RNA segments.
菜豆假单胞菌噬菌体φ6将标记的尿苷三磷酸(UTP)掺入酸不溶性沉淀物中。掺入依赖于乙酸锰、三磷酸腺苷(ATP)、三磷酸鸟苷(GTP)、三磷酸胞苷(CTP)的存在以及对噬菌体进行短时间热处理;该反应受到氯化铵(NH₄Cl)的刺激。用¹⁴C标记的ATP、CTP或GTP替代UTP,并加入适当的未标记核糖核苷三磷酸,结果表明胞苷一磷酸(CMP)掺入量最大,其次是鸟苷一磷酸(GMP)、尿苷一磷酸(UMP)和腺苷一磷酸(AMP)。反应形成的放射性核糖核酸(RNA)在高盐条件下对核糖核酸酶A和T₁具有抗性,但在低盐条件下对这些核酸酶敏感。标记的RNA与φ6双链(ds)RNA一起沉降和共电泳。然而,根据反应中使用的¹⁴C标记核糖核苷三磷酸的不同,放射性在三种ds - RNA组分中的分布有所不同。UMP主要掺入两个较小的ds - RNA片段,GMP主要掺入较大的ds - RNA片段,CMP和AMP在所有三个ds - RNA片段中的分布大致相等。