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首页> 外文期刊>Journal of Virology >Use of lambda gt11 to isolate genes for two pseudorabies virus glycoproteins with homology to herpes simplex virus and varicella-zoster virus glycoproteins.
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Use of lambda gt11 to isolate genes for two pseudorabies virus glycoproteins with homology to herpes simplex virus and varicella-zoster virus glycoproteins.

机译：使用Lambda GT11与同源性与单纯疱疹病毒和毒液疱疹病毒糖蛋白的同源性分离出两个伪病毒糖蛋白的基因。

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A library of pseudorabies virus (PRV) DNA fragments was constructed in the expression cloning vector lambda gt11. The library was screened with antisera which reacted with mixtures of PRV proteins to isolate recombinant bacteriophages expressing PRV proteins. By the nature of the lambda gt11 vector, the cloned proteins were expressed in Escherichia coli as beta-galactosidase fusion proteins. The fusion proteins from 35 of these phages were purified and injected into mice to raise antisera. The antisera were screened by several different assays, including immunoprecipitation of [14C]glucosamine-labeled PRV proteins. This method identified phages expressing three different PRV glycoproteins: the secreted glycoprotein, gX; gI; and a glycoprotein that had not been previously identified, which we designate gp63. The gp63 and gI genes map adjacent to each other in the small unique region of the PRV genome. The DNA sequence was determined for the region of the genome encoding gp63 and gI. It was found that gp63 has a region of homology with a herpes simplex virus type 1 (HSV-1) protein, encoded by US7, and also with varicella-zoster virus (VZV) gpIV. The gI protein sequence has a region of homology with HSV-1 gE and VZV gpI. It is concluded that PRV, HSV, and VZV all have a cluster of homologous glycoprotein genes in the small unique components of their genomes and that the organization of these genes is conserved.
机译：在表达克隆载体Lambda gt11中构建了伪论病毒（PRV）DNA片段的库。用抗血清筛选文库，其与PRV蛋白的混合物反应，以分离表达PRV蛋白的重组噬菌体。通过Lambda GT11载体的性质，克隆蛋白质在大肠杆菌中表达为β-半乳糖苷酶融合蛋白。纯化来自35个噬菌体的融合蛋白并注射到小鼠中以提高抗血清。抗血清被几种不同的测定筛选，包括[14C]葡萄糖胺标记的PRV蛋白的免疫沉淀。该方法鉴定了表达三种不同的PRV糖蛋白的噬菌体：分泌的糖蛋白，GX; GI;和先前未识别的糖蛋白，我们指定GP63。在PRV基因组的小独特区域中彼此相邻的GP63和GI基因图。测定DNA序列用于编码GP63和GI的基因组的区域。发现GP63具有与US7编码的单纯疱疹病毒类型1（HSV-1）蛋白质的同源性的同源性，也具有VariCella-Zoster病毒（VZV）GPIV。 GI蛋白序列具有与HSV-1Ge和VZV GPI的同源性的区域。结论是，PRV，HSV和VZV都在其基因组的小独特组分中具有同源糖蛋白基因的簇簇，并且这些基因的组织被保守。

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