EQUINE HERPESVIRUS TYPE-1 DEFECTIVE-INTERFERING (DI) PARTICLE DNA-STRUCTURE - THE CENTRAL REGION OF THE INVERTED REPEAT IS DELETED FROM DI DNA
EQUINE HERPESVIRUS TYPE-1 DEFECTIVE-INTERFERING (DI) PARTICLE DNA-STRUCTURE - THE CENTRAL REGION OF THE INVERTED REPEAT IS DELETED FROM DI DNA
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DOI：
10.1016/0042-6822(87)90356-4
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发表时间：
1987-07-01
期刊：
影响因子：
3.7
通讯作者：
OCALLAGHAN, DJ
中科院分区：
文献类型：
--
作者：
BAUMANN, RP;STACZEK, J;OCALLAGHAN, DJ
马疱疹病毒1型（EHV-1）基因组的倒置重复序列（IRs）成分是一个重要的结构和功能区域。它是EHV-1缺陷干扰（DI）颗粒DNA的主要成分，该颗粒介导了仓鼠胚胎细胞的致癌转化和持续感染的共同建立。此外，IRs编码单个EHV-1即时早期基因和31.5K极早期蛋白。将包含EHV-1内部IRs和长(L)区与短(S)区连接的DNA序列亚克隆到质粒载体pBR322和pUC12中。共获得22个亚克隆，包括pBR322中的6个Sal亚克隆和pUC12中的12个Sma 1亚克隆。个体亚克隆采用Southern blot杂交来确定亚克隆与EHV-1基因组中重复的、独特的或异质的（het） DNA序列的同源性。这些研究表明，EHV-1热区完全包含在病毒基因组独特的长区域内，并且与L/S连接处相隔约1.8 MDa的完全独特的DNA序列。此外，利用这些IRs亚克隆进行印迹杂交，分析了EHV-1/DI颗粒克隆DNA中IRs DNA序列的完整性。这些分析表明，在DI DNA中存在的IRs DNA序列被广泛重排，并包含主要缺失（0.80-0.83个图谱单位），这使得位于IRs的单个EHV-1立即早期基因（0.78-0.83和0.95-1.00个图谱单位）的大部分被删除。因此，这些数据和之前的研究（R.P. Bauman et al., 1984， J. Virol. 50,13 -21； R.P. Baumann， J. Staczek, and D. J. O-Callaghan, 1986, Virology 153, 188-200）表明，DI DNA分子的主要亚基由IRs成分的精选序列和位于标准病毒基因组L区末端的高度保守的短序列组成。
The inverted repeat (IRs) component of the genome of equine herpesvirus type 1 (EHV-1) is an important region of structure and function. It is a major constituent of the DNA of EHV-1 defective-interfering (DI) particles which have been shown to mediate the coestablishment of oncogenic transformation and persistent infection of hamster embryo cells. In addition, the IRs encodes the single EHV-1 immediate early gene and the 31.5K very early protein. DNA sequences encompassing EHV-1 internal IRs and the joint between the long (L) and short (S) regions were subcloned into the plasmid vectors pBR322 and pUC12. A total of 22 subclones were derived, including six Sal l subclones in pBR322 and 12 Sma l subclones in pUC12. Individual subclones were employed in Southern blot hybridizations to define subclone homology to repeated, unique, or heterogeneous (het) DNA sequences within the EHV-1 genome. These studies revealed that the EHV-1 het region is contained entirely within the unique long region of the viral genome and is separated from the L/S junction by approximately 1.8 MDa of completely unique DNA sequences. Furthermore, these IRs subclones were employed in blot hybridizations to analyze the integrity of IRs DNA sequences within the cloned DNA of EHV-1/DI particles. These analyses demonstrated that IRs DNA sequences present in DI DNA were extensively rearranged and contained major deletions (0.80-0.83 map units) which removed a large portion of the single EHV-1 immediate early gene (0.78-0.83 and 0.95-1.00 map units) located in the IRs. Thus, these data and those previous studies (R.P. Bauman et al., 1984, J. Virol. 50, 13-21; R. P. Baumann, J. Staczek, and D. J. O-Callaghan, 1986, Virology 153, 188-200) indicate that the major subunits of the DI DNA molecule are comprised of selected sequences from the IRs component and a highly conserved short sequence located at the terminus of the L region of the standard viral genome.