CAJ | 학술논문

本研究运用同源重组的方法，在伪狂犬病毒（Pseudorabies virus, PRV）变异株 PRV JS-2012基础上，构建了一株携带增强型绿色荧光蛋白基因的标记重组伪狂犬病毒，命名为rPRV JS-2012-EGFP。经PCR方法鉴定及荧光显微镜观察分析，结果表明重组病毒构建成功，并能在感染细胞中稳定表达绿色荧光蛋白。生长曲线和空斑试验结果显示，rPRV JS-2012-EGFP与亲本株 PRV JS-2012在体外具有相似的生长特性。将 rPRV JS-2012-EGFP毒接种14日龄PRV阴性仔猪，能使实验猪100%发病，病死率高达40%，表明rPRV JS-2012-EGFP是一株具有强致病力的标记伪狂犬病毒变异株，在伪狂犬病毒变异株致病机制研究中具有一定的使用价值。
본연구운용동원중조적방법，재위광견병독（Pseudorabies virus, PRV）변이주 PRV JS-2012기출상，구건료일주휴대증강형록색형광단백기인적표기중조위광견병독，명명위rPRV JS-2012-EGFP。경PCR방법감정급형광현미경관찰분석，결과표명중조병독구건성공，병능재감염세포중은정표체록색형광단백。생장곡선화공반시험결과현시，rPRV JS-2012-EGFP여친본주 PRV JS-2012재체외구유상사적생장특성。장 rPRV JS-2012-EGFP독접충14일령PRV음성자저，능사실험저100%발병，병사솔고체40%，표명rPRV JS-2012-EGFP시일주구유강치병력적표기위광견병독변이주，재위광견병독변이주치병궤제연구중구유일정적사용개치。
An enhanced green fluorescent protein (EGFP) expression cassette was homologously recombined into the genome of a virulent Pseudorabies virus (PRV) variant strain PRV JS-2012, generating a recombinant PRV strain rPRV JS-2012-EGFP. The insert of EGFP gene were confirmed by PCR, and green fluorescence proteins were stably expressed in the cultured cells infected with rPRV JS-2012-EGFP by green fluorescence microscopy analysis. The assays of growth curve and plaque morphology revealed that rPRV JS-2012-EGFP showed similar virological properties to its parent strain PRV JS-2012. 14-day old piglets free of PRV antibodies were inoculated with rPRV JS-2012-EGFP. The morbidity of the infected piglets reached 100%, and the mortality reached 40%. These results showed that rPRV JS-2012-EGFP was a virulent marker PRV and could be utilized to study pathogenic characterization of PRV variant.