Wittek R, Richner B, Hiller G
Nucleic Acids Res. 1984 Jun 25;12(12):4835-48. doi: 10.1093/nar/12.12.4835.
We have mapped the genes coding for two major structural polypeptides of the vaccinia virus core by hybrid selection and transcriptional mapping. First, RNA was selected by hybridization to restriction fragments of the vaccinia virus genome, translated in vitro and the products were immunoprecipitated with antibodies against the two polypeptides. This approach allowed us to map the genes to the left hand end of the largest Hind III restriction fragment of 50 kilobase pairs. Second, transcriptional mapping of this region of the genome revealed the presence of the two expected RNAs. Both RNAs are transcribed from the leftward reading strand and the 5'-ends of the genes are separated by about 7.5 kilobase pairs of DNA. Thus, two genes encoding structural polypeptides with a similar location in the vaccinia virus particle are clustered at approximately 105 kilobase pairs from the left hand end of the 180 kilobase pair vaccinia virus genome.
我们通过杂交筛选和转录图谱分析,确定了编码痘苗病毒核心两种主要结构多肽的基因。首先,通过与痘苗病毒基因组的限制性片段杂交来选择RNA,在体外进行翻译,然后用针对这两种多肽的抗体对产物进行免疫沉淀。这种方法使我们能够将基因定位到50千碱基对的最大Hind III限制性片段的左端。其次,对基因组的这一区域进行转录图谱分析,发现了两种预期的RNA。这两种RNA均从向左转录的链转录而来,基因的5′端被约7.5千碱基对的DNA隔开。因此,在痘苗病毒颗粒中位置相似的两种编码结构多肽的基因,聚集在距180千碱基对的痘苗病毒基因组左端约105千碱基对处。