Meulenberg J J, Bos-de Ruijter J N, Wensvoort G, Moormann R J
Institute for Animal Science and Health, Lelystad, The Netherlands.
Adv Exp Med Biol. 1998;440:199-206. doi: 10.1007/978-1-4615-5331-1_24.
A plasmid containing a full-length cDNA copy of the Lelystad virus isolate (LV) of porcine reproductive and respiratory syndrome virus was constructed. When RNA that was transcribed in vitro from this full-length cDNA clone was transfected to BHK-21 cells, infectious LV was produced and secreted. The virus was rescued by passage to porcine alveolar lung macrophages or CL2621 cells. When infectious transcripts were transfected to porcine alveolar lung macrophages or CL2621 cells no infectious virus was produced due to the poor transfection efficiency of these cells. The growth properties of the viruses produced by BHK-21 cells transfected with infectious transcripts of LV cDNA resembled the growth properties of the parental virus from which the cDNA was derived. The infectious clone of LV enables us to mutagenize the viral genome at specific sites and thus will be useful for detailed molecular characterization of the virus, as well as for the development of a safe and effective live vaccine for use in pigs.
构建了一个包含猪繁殖与呼吸综合征病毒莱利斯塔德病毒分离株（LV）全长cDNA拷贝的质粒。当从该全长cDNA克隆体外转录的RNA转染至BHK-21细胞时，产生并分泌了具有感染性的LV。通过传代至猪肺泡巨噬细胞或CL2621细胞拯救了该病毒。当将感染性转录本转染至猪肺泡巨噬细胞或CL2621细胞时，由于这些细胞的转染效率低，未产生感染性病毒。用LV cDNA感染性转录本转染的BHK-21细胞产生的病毒的生长特性类似于衍生出该cDNA的亲本病毒的生长特性。LV的感染性克隆使我们能够在特定位点诱变病毒基因组，因此将有助于对该病毒进行详细的分子特征分析，以及开发用于猪群的安全有效的活疫苗。
