Physical location of a herpes simplex virus type-1 gene function(s) specifically associated with a 10 million-fold increase in HSV neurovirulence.
Physical location of a herpes simplex virus type-1 gene function(s) specifically associated with a 10 million-fold increase in HSV neurovirulence.
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单纯疱疹病毒 1 型基因功能的物理位置与 HSV 神经毒力增加 1000 万倍特别相关。
DOI：
10.1016/0042-6822(83)90544-5
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发表时间：
1983
期刊：
影响因子：
3.7
通讯作者：
Stevens,JG
中科院分区：
文献类型：
--
作者：
Thompson,RL;Wagner,EK;Stevens,JG
在本文中，我们提出了一种技术，以物理定位HSV-1基因，恢复神经毒力到非神经毒的HSV型间重组在前面的报告中描述。简而言之，组织培养细胞共转染单位长度RE6 DNA和限制性内切酶片段HSV-1（菌株17 Syn+） DNA。这样就产生了RE6和17 Syn+之间的随机重组。然后在小鼠大脑中进行Anin体内富集，以选择含有与神经毒力相关的HSV-1基因的重组体。在用这种方法分离神经毒性重组的5例病例中，限制性内切酶和南部DNA转移分析显示，从0.71到0.83个图谱单元的HSV-1信息已被纳入RE6基因组。通过电泳纯化的HSV-1 DNA片段（包含该区域）进行类似的共转染和体内拯救实验，证实了这部分基因组对HSV神经毒力的作用。随后对该程序生成的神经毒性重组基因的基因组结构分析显示，RE6中仅包含0.71 ~ 0.83个图谱单元的1型信息。因此，存在于病毒DNA这一区域的HSV-1基因功能与病毒的神经毒性增加1000万倍有关。讨论了该技术在活体选择中的潜在应用。
In this paper we present a technique developed to physically locate the HSV-1 gene(s) which restore neurovirulence to a non-neurovirulent HSV intertypic recombinant described in the preceding report. In brief, tissue culture cells are co-transfected with unit length RE6 DNA and restriction endonuclease fragmented HSV-1 (strain 17 Syn+) DNA. In this way, random recombinations between RE6 and 17 Syn+are produced. Anin vivoenrichment in mouse brains is then employed to select recombinants which have incorporated the HSV-1 gene(s) associated with neurovirulence. In each of five cases where neurovirulent recombinants were isolated by this procedure, restriction enzyme and Southern DNA transfer analysis revealed that HSV-1 information from 0.71 to 0.83 map units had been incorporated into the RE6 genome. Confirmation of the role of this portion of the genome for HSV neurovirulence was obtained by similar cotransfection andin vivorescue experiments performed with an electrophoretically purified HSV-1 DNA fragment which encompasses this region. Subsequent genome structure analysis of neurovirulent recombinants generated by this procedure revealed that only type-1 information from 0.71 to 0.83 map units had been incorporated into RE6. Thus an HSV-1 gene function(s) which resides in this region of the viral DNA is associated with a 10 million-fold increase in the neurovirulence of the virus. Potential applications of thisin vivoselection technique are discussed.
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