Lai M M, Hu S S, Vogt P K
Proc Natl Acad Sci U S A. 1977 Nov;74(11):4781-5. doi: 10.1073/pnas.74.11.4781.
The genome size of 20 transformation-defective (td) viruses derived from different strains of Rous sarcoma viruses [Prague (subgroups A and C), Schmidt-Ruppin (subgroups A and D) (SR-D), Bratislava 77, and Carr-Zilber subgroup D)] was examined by polyacrylamide gel electrophoresis. All of the td viruses except td SR-D have 35S RNA of the same size-i.e., class b RNA. Two of five td SR-D viruses examined have a slightly larger RNA, corresponding to a td deletion that is about 25% smaller than that of class b RNA. However, the RNase T(1)-oligonucleotide fingerprints of all the td SR-D viruses are identical, lacking two sarcoma-specific oligonucleotides. The fingerprints of these viruses also showed a minor oligonucleotide present at very low concentration. A study of heteroduplex molecules formed between genome-length cDNA made from wild-type SR-D and 35S RNA of td SR-D showed a deletion loop of 2.0 and 1.5 kilobases, respectively, at the map position of the src gene for these two classes of td SR-D viruses, confirming the results of polyacrylamide gel electrophoresis. In addition, some heteroduplex molecules with a substitution loop of 0.6-0.7 kilobase at the same site as the deletion loop were observed in all five of the td SR-D viruses. We conclude that some of the td SR-D viruses have a partially deleted src gene and that all of the td SR-D viruses have incorporated heterologous sequences of distinct length in some RNA molecules at the position of the src gene. The nature and origin of these heterologous sequences are discussed.
通过聚丙烯酰胺凝胶电泳检测了来自劳氏肉瘤病毒不同毒株(布拉格株(A和C亚组)、施密特 - 鲁平株(A和D亚组)(SR - D)、布拉迪斯拉发77株以及卡尔 - 齐尔伯D亚组)的20种转化缺陷型(td)病毒的基因组大小。除td SR - D外,所有td病毒都具有相同大小的35S RNA,即b类RNA。所检测的5种td SR - D病毒中有2种具有稍大的RNA,对应于一个比b类RNA缺失约小25%的td缺失。然而,所有td SR - D病毒的核糖核酸酶T(1) - 寡核苷酸指纹图谱是相同的,缺少两个肉瘤特异性寡核苷酸。这些病毒的指纹图谱还显示存在一种浓度极低的次要寡核苷酸。对由野生型SR - D制备的基因组长度cDNA与td SR - D的35S RNA形成的异源双链分子的研究表明,这两类td SR - D病毒在src基因的图谱位置分别有2.0和1.5千碱基的缺失环,证实了聚丙烯酰胺凝胶电泳的结果。此外,在所有5种td SR - D病毒中均观察到一些在与缺失环相同位置有0.6 - 0.7千碱基替代环的异源双链分子。我们得出结论,一些td SR - D病毒具有部分缺失的src基因,并且所有td SR - D病毒在src基因位置的一些RNA分子中都掺入了不同长度的异源序列。讨论了这些异源序列的性质和来源。