Kirisawa R, Ohmori H, Iwai H, Kawakami Y
Department of Veterinary Microbiology, Rakuno Gakuen University, Hokkaido, Japan.
Arch Virol. 1993;129(1-4):11-22. doi: 10.1007/BF01316881.
The DNAs from nine Japanese field isolates of equine herpesvirus-1 (EHV-1) were analyzed by digestion with the restriction endonuclease Bam HI and Southern hybridization. Comparing restriction profiles among the EHV-1 strains, there was no considerable difference between isolates before and after vaccine application, but some minor variations in the mobility of Bam HI fragments were observed. To identify these variable fragments, all genomic DNA sequences of the Japanese prototype of EHV-1 have been cloned as Bam HI restriction fragments into the plasmid pUC-18. Physical maps of the virus DNA were constructed by a combination of Southern blot analysis and double enzyme digestion of the cloned fragments. By using these cloned fragments as probes in Southern blot analysis, the areas of heterogeneity observed among the field EHV-1 isolates were located in both terminals of UL, the center of UL, IR, US and TR regions of the genome.
用限制性内切酶Bam HI消化并进行Southern杂交,对来自9株日本马疱疹病毒1型(EHV-1)田间分离株的DNA进行了分析。比较EHV-1毒株之间的限制性图谱,疫苗应用前后的分离株之间没有显著差异,但观察到Bam HI片段迁移率有一些微小变化。为了鉴定这些可变片段,已将EHV-1日本原型株的所有基因组DNA序列作为Bam HI限制性片段克隆到质粒pUC-18中。通过Southern印迹分析和对克隆片段的双酶切相结合构建了病毒DNA的物理图谱。通过在Southern印迹分析中使用这些克隆片段作为探针,在田间EHV-1分离株中观察到的异质性区域位于UL的两个末端、UL的中心、基因组的IR、US和TR区域。