Identification and characterization of a porcine monocytic cell line supporting porcine reproductive and respiratory syndrome virus (PRRSV) replication and progeny virion production by using an improved DNA-launched PRRSV reverse genetics system
Identification and characterization of a porcine monocytic cell line supporting porcine reproductive and respiratory syndrome virus (PRRSV) replication and progeny virion production by using an improved DNA-launched PRRSV reverse genetics system
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DOI：
10.1016/j.virusres.2009.05.013
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发表时间：
2009-10-01
期刊：
影响因子：
5
通讯作者：
Meng, X. J.
中科院分区：
文献类型：
--
作者：
Huang, Y. W.;Fang, Y.;Meng, X. J.
在这项研究中，一个改进的DNA发射（质粒DNA转染为基础）的反向遗传学系统，以降低成本和劳动力的猪繁殖与呼吸综合征病毒（PRRSV）通过引入核酶元件在两个末端的病毒基因组cDNA，置于真核杂交启动子的控制下。通过转染细胞中的报告基因GFP水平和恢复病毒的峰值滴度确定，用该系统拯救PRRSV的效率比基于体外转录RNA的系统和没有工程化核酶元件的传统DNA发射系统高约10-50倍。通过使用这种新的反向遗传学系统，我们确定和特点的猪单核细胞系，3D 4/31，能够支持PRRSV复制，子代病毒粒子的生产，并附着在细胞表面。该反向遗传系统的建立和支持PRRSV复制的猪单核细胞系的鉴定将有助于进一步研究PRRSV的宿主-病毒相互作用。(C)2009 Elsevier B. V.保留所有权利。
In this study, an improved DNA-launched (plasmid DNA transfection-based) reverse genetics system with reduced cost and labor was developed for porcine reproductive and respiratory syndrome virus (PRRSV) by introduction of ribozyme elements at both termini of the viral genomic cDNA that were placed under the control of a eukaryotic hybrid promoter. The rescue efficacy of PRRSV with this system was approximately 10-50-fold higher than the in vitro-transcribed RNA-based system and the traditional DNA-launched system without the engineered ribozyme elements, as determined by reporter GFP level in transfected cells and the peak titer of the recovery virus. By using this new reverse genetics system, we identified and characterized a porcine monocytic cell line, 3D4/31, capable of supporting PRRSV replication, progeny virion production, and attachment on the cell surface. The establishment of this improved reverse genetic system and the identification of a porcine monocytic cell line supporting PRRSV replication will aid future studies of host-virus interaction of PRRSV. (C) 2009 Elsevier B.V. All rights reserved.