STRUCTURE OF THE GENOME OF EQUINE HERPESVIRUS TYPE-1
STRUCTURE OF THE GENOME OF EQUINE HERPESVIRUS TYPE-1
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DOI：
10.1016/0042-6822(81)90092-1
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发表时间：
1981-01-01
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：
HENRY, BE;ROBINSON, RA;OCALLAGHAN, DJ
通过限制性内切酶图谱研究确定了马疱疹病毒1型(EHV-1)基因组的分子结构。对纯化的EHV-1 DNA，无论是未标记的，32PO4标记的，还是[~3H]TdR[脱氧胸腺嘧啶核苷]标记的，一级限制性内切酶的酶切图谱如下：EcoRI产生17个片段，大小为23.4-1.3Md；BglII，16个片段，大小为24.5-1.0md；XbaI，15个主要片段，大小为18.6-1.7md；BamHI，17个大小片段，大小为13.7-2.8md。有几个片段的量为0.5M，其余的均为1.0M；未检测到0.25M的片段。用不同的酶对这些分离的片段进行二级限制性内切酶消化，使用.lambda的两种方法分析末端片段。5‘端核酸外切、多核苷酸激酶末端标记和32P标记的BamHI片段杂交实验表明，该疱疹病毒基因组是一个92md的线性双链DNA分子，由L(长)和S(短)两个片段组成，分别为71.6md和20.4md。0.5M片段位于S区域的末端，这种排列允许S区域相对于L区域反转；因此，基因组存在两种结构排列(异构体)。在L末端、S片段内和L地区的分裂变异位点处检测到了异质性区域。
The molecular structure of the genome of equine herpesvirus type 1 (EHV-1) was determined by restriction endonuclease mapping studies. Primary restriction enzyme digestion of purified EHV-1 DNA, either unlabeled, 32PO4 labeled or [3H]TdR [deoxythymidine] labeled, gave the following cleavage patterns: EcoRI yielded 17 fragments of 23.4-1.3 megadaltons (Md); BglII, 16 fragments of 24.5-1.0 Md; XbaI, 15 major fragments of 18.6-1.7 Md; and BamHI, 17 fragments of 13.7-2.8 Md. Several fragments were present in 0.5 M amounts while all others were 1.0 M; no 0.25 M fragments were detected. Secondary restriction enzyme digestion of these isolated fragments with various enzymes, analysis of terminal fragments using both the methods of .lambda. 5'' exonuclease digestion and end labeling with polynucleotide kinase and blot hybridization experiments with 32P-labeled BamHI fragments indicated that this herpesvirus genome is a 92-Md linear, double-stranded DNA molecule and is comprised of 2 segments designated L (long) and S (short) which are 71.6 and 20.4 Md, respectively. The 0.5 M fragments are located at the ends of the S region, an arrangement which allows the S region to invert relative to the L region; thus, 2 structural arrangements (isomers) of the genome exist. Areas of heterogeneity were detected at the L terminus, within the S segment and at a split variable locus in the L region.