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通过聚合酶链反应(PCR)和限制性内切酶分析区分血清学相关的反刍动物α疱疹病毒。

The distinction of serologically related ruminant alphaherpesviruses by the polymerase chain reaction (PCR) and restriction endonuclease analysis.

作者信息

Lyaku J R, Vilcek S, Nettleton P F, Marsden H S

机构信息

Moredun Research Institute, Edinburgh, UK.

出版信息

Vet Microbiol. 1996 Jan;48(1-2):135-42. doi: 10.1016/0378-1135(95)00136-0.

Abstract

The amplification and analysis of a 468bp fragment from the gB gene of the serologically related ruminant alphaherpesviruses bovine herpesvirus-1.1 and 1.2 (BHV-1.1 and BHV-1.2), caprine herpesvirus-1 (CapHV-1), cervine herpesvirus-1 (CerHV-1) and rangiferine herpesvirus-1 (RanHV-1) by PCR and restriction endonuclease analysis is described. As primers, 22bp oligomers selected from the BHV-1 gB gene sequences were used for the amplification of the DNA from the five viruses. The amplification product from each virus was analysed by the restriction endonuclease enzymes BglI, HinfI, SmaI and AvaI. The specific amplification obtained demonstrate the existence of the gB gene sequences for each of the five alphaherpesviruses. However, sequences from some of the fragments were found to be different from those predicted from the gB gene following restriction endonuclease analysis. All five amplification products generated the same number of fragments after digestion with HinfI except for two additional bands evident in CapHV-1. The CerHV-1 and RanHV-1 fragments contained slightly different BglI restriction sites from those of the other three. While BHV-1.1, BHV-1.2, CapHV-1 and CerHV-1 contained SmaI and AvaI restriction sites, the RanHV-1 amplification product lacked both SmaI and AvaI restriction sites.

摘要

描述了通过聚合酶链反应(PCR)和限制性内切酶分析对血清学相关的反刍动物α疱疹病毒牛疱疹病毒1.1和1.2(BHV - 1.1和BHV - 1.2)、山羊疱疹病毒1(CapHV - 1)、鹿疱疹病毒1(CerHV - 1)和驯鹿疱疹病毒1(RanHV - 1)的gB基因的468bp片段进行扩增和分析。作为引物,从BHV - 1 gB基因序列中选择的22bp寡聚物用于扩增这五种病毒的DNA。来自每种病毒的扩增产物用限制性内切酶BglI、HinfI、SmaI和AvaI进行分析。所获得的特异性扩增证明了这五种α疱疹病毒各自gB基因序列的存在。然而,在限制性内切酶分析后发现,一些片段的序列与从gB基因预测的序列不同。用HinfI消化后,除了CapHV - 1中明显的另外两条带外,所有五种扩增产物产生的片段数量相同。CerHV - 1和RanHV - 1片段的BglI限制性位点与其他三种略有不同。虽然BHV - 1.1、BHV - 1.2、CapHV - 1和CerHV - 1含有SmaI和AvaI限制性位点,但RanHV - 1扩增产物缺乏SmaI和AvaI限制性位点。

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