Bejcek B, Conley A J
Virology. 1986 Oct 15;154(1):41-55. doi: 10.1016/0042-6822(86)90428-9.
Morphologically transformed rat (3Y1) cell lines were established following transfection with HSV-2 mtrII DNA sequences (0.585 to 0.601 map units). The mtrII sequences were cloned in plasmids containing the neor gene. Cells resistant to the antibiotic G-418 were passed into soft agarose, and clonal lines were established from individual colonies. The DNAs from two cell lines examined by Southern blot hybridization were shown to contain the original transfected viral DNA sequences in a fashion consistent with a multiple and complex pattern of integration. From one cell line, an approximately 20-kbp plasmid was isolated after transformation of bacteria with Hirt supernatant DNA. This plasmid was capable of rapidly transforming rat cells at a greater than 1000-fold higher frequency than the mtrII DNA. This plasmid consists mainly of unique sequence rat DNA with two copies of the HSV-2 mtrII region DNA (HSV-2 genomic map unit location of ca. 0.595) present at sites distant from each other. The rat DNA in the rescued plasmid is homologous to the putative focus-forming sequences present in the HSV-2 mtrIII (0.53 to 0.58 map units) and the colinear HSV-1 DNA. The genomic copy of these rat sequences in four HSV-2 mtrII transformed cell lines appears to have undergone rearrangement. These data provide evidence that the HSV-2 mtrII sequences are involved in transformation, and that the HSV-2 mtrII region may affect transformation by rearranging the cellular sequences that are homologous to mtrIII.
用单纯疱疹病毒2型（HSV - 2）mtrII DNA序列（0.585至0.601图谱单位）转染后，建立了形态学转化的大鼠（3Y1）细胞系。mtrII序列克隆于含有新霉素抗性基因（neor）的质粒中。对抗生素G - 418有抗性的细胞被接种到软琼脂糖中，并从单个菌落建立克隆系。通过Southern印迹杂交检测的两个细胞系的DNA显示，其含有原始转染的病毒DNA序列，整合方式呈现多重且复杂的模式。从一个细胞系中，在用Hirt上清液DNA转化细菌后分离出一个约20kbp的质粒。该质粒能够以比mtrII DNA高1000倍以上的频率快速转化大鼠细胞。该质粒主要由独特序列的大鼠DNA组成，在彼此远离的位点存在两个HSV - 2 mtrII区域DNA拷贝（HSV - 2基因组图谱单位位置约为0.595）。拯救质粒中的大鼠DNA与HSV - 2 mtrIII（0.53至0.58图谱单位）和共线性HSV - 1 DNA中存在的假定集落形成序列同源。在四个HSV - 2 mtrII转化细胞系中，这些大鼠序列的基因组拷贝似乎发生了重排。这些数据提供了证据，表明HSV - 2 mtrII序列参与转化，并且HSV - 2 mtrII区域可能通过重排与mtrIII同源的细胞序列来影响转化。
