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在昆虫细胞中表达含有甲型肝炎病毒多蛋白抗原表位的重组轮状病毒蛋白。

Expression of recombinant rotavirus proteins harboring antigenic epitopes of the hepatitis a virus polyprotein in insect cells.

机构信息

Department of Microbiology, Chung-Ang University College of Medicine, Seoul 156-756.

出版信息

Biomol Ther (Seoul). 2012 May;20(3):320-5. doi: 10.4062/biomolther.2012.20.3.320.

Abstract

Rotavirus and hepatitis A virus (HAV) spread by the fecal-oral route and infections are important in public health, especially in developing countries. Here, two antigenic epitopes of the HAV polyprotein, domain 2 (D2) and domain 3 (D3), were recombined with rotavirus VP7, generating D2/VP7 and D3/VP7, cloned in a baculovirus expression system, and expressed in Spodoptera frugiperda 9 (Sf9) insect cells. All were highly expressed, with peak expression 2 days post-infection. Western blotting and ELISA revealed that two chimeric proteins were antigenic, but only D2/VP7 was immunogenic and elicited neutralizing antibody responses against rotavirus and HAV by neutralization assay, implicating D2/VP7 as a multivalent subunit-vaccine Candidate for preventing both rotavirus and HAV infections.

摘要

轮状病毒和甲型肝炎病毒(HAV)通过粪-口途径传播,感染在公共卫生中非常重要,尤其是在发展中国家。在这里,HAV 多蛋白的两个抗原表位,结构域 2(D2)和结构域 3(D3)与轮状病毒 VP7 重组,生成 D2/VP7 和 D3/VP7,克隆在杆状病毒表达系统中,并在 S 型粉纹夜蛾 9( Sf9)昆虫细胞中表达。所有这些都高度表达,感染后 2 天达到峰值表达。Western blotting 和 ELISA 显示两种嵌合蛋白具有抗原性,但只有 D2/VP7 具有免疫原性,并通过中和试验诱导针对轮状病毒和 HAV 的中和抗体反应,表明 D2/VP7 是一种多价亚单位疫苗候选物,可预防轮状病毒和 HAV 感染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7b2a/3794530/2fbe26fae87d/ooomb4-20-320-g001.jpg

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