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TRANSFORMATION INDUCED BY THE MTRII OF HERPES SIMPLEX TYPE 2.

机译:单纯疱疹2型基质引起的转化

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摘要

Herpes simplex viruses types 1 and 2 (HSV-1 and HSV-2) have been implicated as causing tumors in humans. In vitro, both viruses will transform a variety of established and primary cells. There appear to be three specific regions that are capable of inducing transformations, one in HSV-1 (mtrI) and two in HSV-2 (mtrII and mtrIII). None of these regions share homology with each other, and all probably transform by different mechanisms. Morphologically transformed rat cell (3Y1) lines were established following transfection with HSV-2 mtrII DNA sequences. This transformation region was cloned into pSV2neo, allowing for the selection of cells resistant to the antibiotic G418. Resistant cells were plated into soft agarose and the resultant colonies used to establish cell lines. 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 kilobase pair plasmid was isolated after transformation of bacteria with Hirt supernatant DNA. This plasmid was capable of rapidly transforming 3Y1 cells at a 1000-fold higher frequency than the plasmid used to create this cell line and consists mainly of unique sequence rat DNA. Two copies of the HSV-2 mtrII region DNA are also present but at sites distant from each other. The rat DNA is homologous to the putative focus forming sequences present in HSV-2 mtrIII 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 in different fashions. These data provide evidence that the HSV-2 mtrII sequences are involved in HSV-2 transformation and suggest that this region may affect transformation by rearranging the cellular sequences that are homologous to mtrIII.
机译:1型和2型单纯疱疹病毒(HSV-1和HSV-2)已被认为可导致人类肿瘤。在体外,两种病毒都将转化多种已建立的原代细胞。似乎存在三个能够诱导转化的特定区域,一个在HSV-1(mtrI)中,一个在HSV-2(mtrII和mtrIII)中。这些区域中没有一个彼此共享同源性,并且都可能通过不同的机制进行转化。用HSV-2 mtrII DNA序列转染后,建立了形态转化的大鼠细胞(3Y1)系。将该转化区克隆到pSV2neo中,从而选择对抗生素G418有抗性的细胞。将抗性细胞接种到软琼脂糖中,所得菌落用于建立细胞系。通过Southern印迹杂交检查的来自两种细胞系的DNA显示出含有原始转染的病毒DNA序列,其方式与多重和复杂的整合模式一致。用Hirt上清液DNA转化细菌后,从一个细胞系中分离出约20kb的质粒对。该质粒能够以比用于创建该细胞系的质粒高1000倍的频率快速转化3Y1细胞,并且主要由独特序列的大鼠DNA组成。还存在两个拷贝的HSV-2 mtrII区DNA,但位于彼此远离的位点。大鼠DNA与HSV-2 mtrIII和共线HSV-1 DNA中存在的推定焦点形成序列同源。这些大鼠序列在四个HSV-2 mtrII转化的细胞系中的基因组拷贝似乎已经以不同的方式进行了重排。这些数据提供了HSV-2 mtrII序列参与HSV-2转化的证据,并表明该区域可通过重新排列与mtrIII同源的细胞序列来影响转化。

著录项

  • 作者

    BEJCEK, BRUCE EVAN.;

  • 作者单位

    Saint Louis University.;

  • 授予单位 Saint Louis University.;
  • 学科 Biology Molecular.
  • 学位 Ph.D.
  • 年度 1986
  • 页码 96 p.
  • 总页数 96
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分子遗传学;
  • 关键词

  • 入库时间 2022-08-17 11:51:01

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