Reverse genetic manipulation of the overlapping coding regions for structural proteins of the type II porcine reproductive and respiratory syndrome virus
文献类型: 外文期刊
第一作者: Yu, Dandan
作者: Yu, Dandan;Lv, Jian;Sun, Zhi;Zheng, Haihong;Lu, Jiaqi;Yuan, Shishan
作者机构:
关键词: PRRSV;Structural protein;Overlapping coding region;Reverse genetic manipulation;Transcription regulating sequence;TRS
期刊名称:VIROLOGY ( 影响因子:3.616; 五年影响因子:3.967 )
ISSN: 0042-6822
年卷期: 2009 年 383 卷 1 期
页码:
收录情况: SCI
摘要: The overlapping genomic regions coding for structural proteins of porcine reproductive and respiratory syndrome virus (PRRSV) poses problems for molecular dissection of the virus replication process. We constructed five mutant full-length cDNA clones with the overlapping regions unwound and 1 to 3 restriction sites inserted between two adjacent ORFs (ORF1/2, ORF4/5, ORF5/6, ORF 6/7 and ORF7/3' UTR), which generated the recombinant viruses. Our findings demonstrated that 1) the overlapping structural protein ORFs can be physically separated, and is dispensable for virus viability; 2) Such ORF separations did not interrupt the subgenomic RNA synthesis; 3) the plaque morphology, growth kinetics, and antigenicity of these mutant viruses were virtually indistinguishable from those of the parental virus in cultured cells; and 4) these mutant viruses remained genetic stable in vitro. This study lays a foundation for further molecular dissection of PRRSV replication process, and development of genetically tagged vaccines against PRRS. (c) 2008 Elsevier Inc. All rights reserved.
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