Schubert M, Keene J D, Lazzarini R A, Emerson S U
Cell. 1978 Sep;15(1):103-12. doi: 10.1016/0092-8674(78)90086-7.
The 2S RNA synthesized in vitro by the RNA polymerase of a defective interfering (DI) particle of vesicular stomatitis virus was labeled at its 3' terminus with 32P-cytidine 3', 5' bisphosphate and RNA ligase. Analysis of the labeled RNA showed that it was a family of RNAs of different length but all sharing the same 5' terminal sequence. The largest labeled RNA was purified by gel electrophoresis, and the sequence of 41 of its 46 nucleotides was determined by rapid RNA sequencing methods. The assignment of the remaining 5 nucleotides was made on the basis of an analysis of one of the smaller RNAs and published data. A new approach in RNA sequencing based on the identification of 3' terminal nucleotides of rna fragments originally present in the DI product or generated during the ligation reaction confirmed most of the sequence. The complete sequence of this 46 nucleotide long plus-sense RNA is: ppACGAAGACCACAAAACCAGAUAAAAAA UAAAAACCACAAGAGGGUC-OH. This RNA anneals to the RNA of the DI particle from which it was synthesized, indicating that its synthesis is template-specified. At least the first 17 and possibly all of the nucleotides are also complementary to sequences at the 3' end of two other VSV DI particles which were derived independently and whose genomes differ significantly in length. These data suggest a common 3' terminal sequence among all VSV DI particles which contain part of the Lgene region of the parental genome.
水疱性口炎病毒缺陷干扰（DI）颗粒的RNA聚合酶在体外合成的2S RNA，用32P - 胞苷3'，5' - 二磷酸和RNA连接酶在其3'末端进行标记。对标记RNA的分析表明，它是一个不同长度的RNA家族，但都共享相同的5'末端序列。通过凝胶电泳纯化最大的标记RNA，并通过快速RNA测序方法确定其46个核苷酸中的41个的序列。其余5个核苷酸的归属是基于对较小RNA之一的分析和已发表的数据。一种基于鉴定最初存在于DI产物中或在连接反应过程中产生的RNA片段的3'末端核苷酸的RNA测序新方法证实了大部分序列。这个46个核苷酸长的正链RNA的完整序列是：ppACGAAGACCACAAAACCAGAUAAAAAA UAAAAACCACAAGAGGGUC - OH。这种RNA与合成它的DI颗粒的RNA退火，表明其合成是由模板指定的。至少前17个核苷酸，可能所有核苷酸也与另外两个独立衍生且基因组长度差异显著的VSV DI颗粒3'末端的序列互补。这些数据表明，所有包含亲本基因组L基因区域一部分的VSV DI颗粒之间存在共同的3'末端序列。
