Construction of a minigenome rescue system for Newcastle disease virus strain Italien
Construction of a minigenome rescue system for Newcastle disease virus strain Italien
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新城疫病毒Italien小基因组拯救系统的构建。
DOI：
10.1007/s00705-010-0898-3
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发表时间：
2011-04-01
影响因子：
2.7
通讯作者：
Bian, Huijie
中科院分区：
文献类型：
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作者：
Feng, Hao;Wei, Ding;Bian, Huijie
意大利纽卡斯尔病病毒（NDV）是一种强毒株，是一种溶瘤病毒，被认为是一种潜在的抗肿瘤病毒治疗剂。以真核表达质粒pcDNA3.1（+）为基础，构建了表达NDV Italien NP、P和L基因的辅助质粒。构建了由NDV Italien的3'前导区和5'尾随区组成的微型基因组，其侧翼是编码萤火虫荧光素酶的报告基因，以检测三种辅助质粒在病毒基因组复制和转录中的功效。在用三种辅助质粒和微型基因组质粒共转染BSR-T7/5细胞后，通过测定荧光素酶活性来评估微型基因组RNA的复制。在微型基因组拯救系统中，检测报告基因的表达。我们的研究结果表明，三种蛋白质NP，P和L的正确表达，可以组装成一个功能性的核糖核蛋白复合物，有效地指导微型基因组RNA的转录。
Newcastle disease virus (NDV) Italien, a velogenic strain, is an oncolytic virus that is considered to be a potential agent for antitumor viral therapy. We constructed three helper plasmids expressing the NP, P and L genes of NDV Italien based on the eukaryotic expression plasmid pcDNA3.1(+). The minigenome consisting of the 3' leader and 5' trailer regions of NDV Italien flanking a reporter gene encoding firefly luciferase was constructed to examine the efficacy of the three helper plasmids in viral genome replication and transcription. After co-transfection of BSR-T7/5 cells with the three helper plasmids and the minigenome plasmid, replication of minigenome RNA was evaluated by determining luciferase activity. In the minigenome rescue system, expression of the reporter gene was detected. Our results indicate that the three proteins NP, P, and L are correctly expressed and can assemble into a functional ribonucleoprotein complex that effectively directs the transcription of minigenome RNA.