中国兽医科学2011，Vol.41Issue(12)：1223-1227,5.
高致病性猪繁殖与呼吸综合征病毒NSP1蛋白重组腺病毒的构建及其对小鼠的免疫抑制作用
Immunosuppression of NSP1 of highly pathogenic porcine reproductive and respiratory syndrome virus expressed in a recombinant adenovirus in mice
摘要
Abstract
The non-structural protein I（NSP1） gene of the highly pathogenic PRRSV strain SY0608 was amplified by RT-PCR and cloned into pShuttle-CMV plasmid, and confirmed by PCR and sequence analysis. The recombinant plasmid was linearized and transformed into E. coli BJ5183 strain together with adenovirus backbone vector pAdEasy-1. After being linearized with Pine I , the recombinant adenovirus plasmid pAd-NSP1 was transferred into HEK293-A cells and the recombinant adenovirus rAd-NSP1 was obtained. The results of Western-blot and IFA showed that the NSP1 could be expressed in 293 cells line infected with the recombinant adenovirus. Forty-five ICR mice were inoculated with rAd-NSP1 subcutaneously twice at 2-week apart, and PRRSV-specific cell mediate immune responses were detected by T lymphocyte proliferation assay and ELISA for IFN-γ and IL-10. The results showed that the mice inoculated with rAd-NSP1 had significantly lower levels of lymphocyte proliferation and IFN-γ, but had obviously higher levels of IL-10 compared with the control group inoculated with wild type adenovirus or medium alone. It indicated that NSP1 of highly pathogenic PRRSV could inhibit the immunity in mice.关键词
高致病性猪繁殖与呼吸综合征病毒/NSP1/重组腺病毒/IFN-γKey words
high pathogenic PRRSV/NSP1/recombinant adenovirus/IFN-γ分类
农业科技引用本文复制引用
周业飞,李玉峰,王先炜,姜平..高致病性猪繁殖与呼吸综合征病毒NSP1蛋白重组腺病毒的构建及其对小鼠的免疫抑制作用[J].中国兽医科学,2011,41(12):1223-1227,5.基金项目
国家自然科学基金项目(30871868) （30871868）
国家重大转基因专项(2009ZX08009-143B) （2009ZX08009-143B）
农业部公益性行业专项子项目(201003060-04) （201003060-04）
国家生猪产业体系建设专项(CARS-36) （CARS-36）
