构建和特性的四环素调控的表达立即早期基因的重组伪狂犬病病毒。

Construction and properties of a recombinant pseudorabies virus with tetracycline-regulated control of immediate-early gene expression.

机构信息

Departamento de Medicina Preventiva, Salud Pública y Microbiología, Facultad de Medicina, Universidad Autónoma de Madrid, Madrid, Spain.

出版信息

J Virol Methods. 2011 Jan;171(1):253-9. doi: 10.1016/j.jviromet.2010.11.011. Epub 2010 Nov 17.

DOI:10.1016/j.jviromet.2010.11.011

PMID:21093486

Abstract

A study was carried out to determine whether altering the control of expression of the IE180 gene of pseudorabies virus (PRV), by replacing the IE180 promoter with the tetracycline-responsive promoter (Ptet), affects virus replication and virulence. This PRV-BT90 mutant virus was constructed by complementation and recombination in Hela Tet-Off cells. The virus yield produced by infection of Hela Tet-Off cells with PRV-BT90 was similar to that of the parental virus vBecker2. Viral replication of PRV-BT90 was reduced in Vero cells as reflected by a reduction of virus yield and plating efficiency compared to vBecker2. PRV-BT90 plaque formation in Hela Tet-Off cells was inhibited in the presence of doxycycline, whereas vBecker2 plaque formation was not affected. Subcutaneous infection of mice with the two viruses revealed a LD(50) higher than 10(6) TCID(50) for the PRV-BT90 mutant virus while the LD(50) was 178 TCID(50) for the vBecker2 parental virus.

摘要

一项研究旨在确定改变伪狂犬病病毒（PRV）IE180 基因的表达控制，通过用四环素反应性启动子（Ptet）替代 IE180 启动子，是否会影响病毒复制和毒力。该 PRV-BT90 突变病毒是通过在 Hela Tet-Off 细胞中互补和重组构建的。用 PRV-BT90 感染 Hela Tet-Off 细胞产生的病毒产量与亲本病毒 vBecker2 相似。PRV-BT90 在 Vero 细胞中的病毒复制减少，表现在病毒产量和噬斑形成效率与 vBecker2 相比降低。在存在强力霉素的情况下，PRV-BT90 在 Hela Tet-Off 细胞中的斑块形成被抑制，而 vBecker2 斑块形成不受影响。用两种病毒对小鼠进行皮下感染，结果表明 PRV-BT90 突变病毒的 LD（50）高于 10（6）TCID（50），而 vBecker2 亲本病毒的 LD（50）为 178 TCID（50）。