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通过限制性内切酶分析和杂交对伪狂犬病(奥耶斯基氏病)病毒野外分离株的评估

Evaluation of field isolates of pseudorabies (Aujeszky's disease) virus as determined by restriction endonuclease analysis and hybridization.

作者信息

Pirtle E C, Wathen M W, Paul P S, Mengeling W L, Sacks J M

出版信息

Am J Vet Res. 1984 Oct;45(10):1906-12.

PMID:6093637
Abstract

Sixty-seven isolates of pseudorabies virus (PRV) from 13 states were cleaved with 4 restriction endonucleases (RE), and after electrophoresis in agarose, their banding patterns were photographed and evaluated. The deoxyribonucleic acid (DNA) cleavage fragments were designated into regions specified by molecular weight ranges based on lambda phage DNA as a size marker. The 67 PRV isolates were evaluated according to the total number of cleavage fragments, by the number of fragments within designated molecular weight regions, and finally, by the migration of fragments within regions. Four of the 67 PRV isolates (all 4 from California) did not have a 4.1 to 4.6 megadalton HinfI band, but hybridization of the HinfI digests with a 32P probe made with the 4.4 megadalton band hybridized with 2 lighter fragments, 2.5 and 1.9 megadaltons, respectively. The BamHI digests of DNA from some PRV isolates with submolar fragments were hybridized with 32P probes made with fragments from the submolar region and the BamHI E fragment. Both probes hybridized to the submolar region of PRV with BamHI submolar fragments, but only to the trimolar (E, F, and G) band of PRV without submolar fragments in the 4.1 to 7.5 BamHI megadalton region. Epidemiologic evidence was obtained which indicated that a Missouri strain of PRV was transferred to an Illinois swine herd by importation of feeder pigs from Missouri. The results indicate that there are numerous genomically different PRV currently in circulation in the United States and that the combination of RE analysis and DNA hybridization offers useful epidemiologic information to evaluate the various strains.

摘要

对来自13个州的67株伪狂犬病病毒(PRV)分离株用4种限制性内切酶(RE)进行切割,在琼脂糖凝胶中电泳后,拍摄并评估其条带模式。以λ噬菌体DNA作为大小标记,将脱氧核糖核酸(DNA)切割片段按分子量范围指定的区域进行划分。根据切割片段的总数、指定分子量区域内的片段数量,最后根据区域内片段的迁移情况对67株PRV分离株进行评估。67株PRV分离株中有4株(均来自加利福尼亚州)没有4.1至4.6兆道尔顿的HinfI条带,但HinfI酶切产物与用4.4兆道尔顿条带制备的32P探针杂交后,与2条较浅的片段杂交,分别为2.5和1.9兆道尔顿。一些具有亚摩尔片段的PRV分离株的DNA的BamHI酶切产物与用亚摩尔区域片段和BamHI E片段制备的32P探针杂交。两种探针均与具有BamHI亚摩尔片段的PRV亚摩尔区域杂交,但仅与在4.1至7.5 BamHI兆道尔顿区域没有亚摩尔片段的PRV的三摩尔(E、F和G)条带杂交。获得的流行病学证据表明,一株密苏里州的PRV毒株通过从密苏里州进口育肥猪转移到了伊利诺伊州的一个猪群中。结果表明,目前美国有许多基因组不同的PRV在传播,并且限制性内切酶分析和DNA杂交相结合可为评估各种毒株提供有用的流行病学信息。

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