Khoury G, Carter B J, Ferdinand F J, Howley P M, Brown M, Martin M A
J Virol. 1976 Mar;17(3):832-40. doi: 10.1128/JVI.17.3.832-840.1976.
The topographical locations on the simian virus 40 (SV40) genome of the templates for virus-specific RNA species present late in the lytic infection were determined by RNA-DNA hybridization experiments with the Hind restriction enzyme fragments. Two classes of late virus-specific cytoplasmic mRNA's can be separated on the basis of either sedimentation properties in neutral sucrose or electrophoretic mobility in polyacrylamide gels. In the 16S class, two species of RNA were identified by hybridization experiments. One of these species was complementary to sequences of the early template on the minus (E) strand (0.175 to 0.655 map units), and the other more abundant species was complementary to sequences present in the late template on the plus (L) strand (0.655 to 0.175 map units). In addition two species were detected in the 16S class of late cytoplasmic virus-specific mRNA. One of these species was the major late RNA detected and consisted of a polyadenylated transcript complementary to the plus (L) DNA strands of Hind fragments K, F, J, and G (0.945 to 0.175 map units). This species appears to specify the major capsid protein (VP1). A less abundant nonpolyadenylated 16S RNA species complementary to the plus (L) strands of Hind fragments C, D, and E (0.655 to 0.945 map units) may result from post-transcriptional processing or nonspecific degradation of the 19S viral RNA complementary to the plus (L) strand.
通过用Hind限制性内切酶片段进行RNA-DNA杂交实验,确定了在猿猴病毒40(SV40)基因组上,在裂解感染后期出现的病毒特异性RNA种类模板的拓扑位置。根据在中性蔗糖中的沉降特性或在聚丙烯酰胺凝胶中的电泳迁移率,可以分离出两类晚期病毒特异性细胞质mRNA。在16S类别中,通过杂交实验鉴定出两种RNA。其中一种与负(E)链上早期模板的序列互补(0.175至0.655图谱单位),另一种更丰富的种类与正(L)链上晚期模板中存在的序列互补(0.655至0.175图谱单位)。此外,在晚期细胞质病毒特异性mRNA的16S类别中检测到两种。其中一种是检测到的主要晚期RNA,由与Hind片段K、F、J和G的正(L)DNA链互补的多聚腺苷酸化转录本组成(0.945至0.175图谱单位)。这种种类似乎指定了主要衣壳蛋白(VP1)。一种与Hind片段C、D和E的正(L)链互补的丰度较低的非多聚腺苷酸化16S RNA种类(0.655至0.945图谱单位)可能是由与正(L)链互补的19S病毒RNA的转录后加工或非特异性降解产生的。