Zhu Yanxin, Mou Qiuju, Zhao Bingbing, Luo Hong, Zhi Yan, Lan Lusha, Jiang Yan
Department of Microbiology, Guizhou Medical University, Guiyang 550004, China.
Experimental Animal Center, Guizhou Medical University, Guiyang 550004, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2015 Dec;31(12):1615-9.
To obtain non-structural protein 2 (NS2) of influenza A virus (IAV) via the expression system of Escherichia coli, and observe the effect of NS2 on the production of interferon γ (IFN-γ) in mouse lung tissues.
A prokaryotic expression plasmid pET22b(+)/NS2 was constructed and transformed into Rosetta-gami(2) of Escherichia coli. Through the inducible expression and purification, NS2 was prepared. Forty BALB/c mice were divided randomly into 8 groups, normal control group, 200 μg/kg NS2 joint virus group, 200 μg/kg NS2 joint RNA group, 100 μg/kg inactivated virus group, 0.2×lethal dose 50% (LD50) virus group, viral RNA group of 50 μg/kg, 200 μg/kg NS2 group, and 100 μg/kg NS2 group. All mice were subjected to nasal inhalation. Reverse transcription-PCR and Western blotting were respectively performed to detect the mRNA and protein expressions of IFN-γ in lung tissues.
Recombinant NS2 was obtained by the induction of isopropyl-beta-D-thiogalactopyranoside (IPTG) and purification. After the nasal inhalation of NS2 or/and IAV to mice, reverse transcription-PCR and Western blotting showed that the levels of IFN-γ mRNA and protein in the lung tissues of NS2 joint virus group were lower than those of the virus group.
NS2 could inhibit the expressions of IFN-γ mRNA and protein in lung tissues of mice infected by IAV.
通过大肠杆菌表达系统获得甲型流感病毒（IAV）的非结构蛋白2（NS2），并观察NS2对小鼠肺组织中干扰素γ（IFN-γ）产生的影响。
构建原核表达质粒pET22b(+)/NS2，并将其转化至大肠杆菌Rosetta-gami(2)中。通过诱导表达和纯化制备NS2。将40只BALB/c小鼠随机分为8组，即正常对照组、200 μg/kg NS2联合病毒组、200 μg/kg NS2联合RNA组、100 μg/kg灭活病毒组、0.2×半数致死量（LD50）病毒组、50 μg/kg病毒RNA组、200 μg/kg NS2组和100 μg/kg NS2组。所有小鼠均进行滴鼻给药。分别采用逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测肺组织中IFN-γ的mRNA和蛋白表达。
经异丙基-β-D-硫代半乳糖苷（IPTG）诱导和纯化获得重组NS2。对小鼠滴鼻给予NS2或/和IAV后，RT-PCR和蛋白质免疫印迹法检测显示，NS2联合病毒组肺组织中IFN-γ的mRNA和蛋白水平低于病毒组。
NS2可抑制IAV感染小鼠肺组织中IFN-γ的mRNA和蛋白表达。
