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使用重组杆状病毒对马疱疹病毒1型糖蛋白H进行表达及特性分析

Expression and characterisation of equine herpesvirus 1 glycoprotein H using a recombinant baculovirus.

作者信息

McGowan E M, Hayden M B, Edwards S J, Pye D, Love D N, Whalley J M

机构信息

School of Biological Sciences, Macquarie University, Sydney, New South Wales, Australia.

出版信息

Arch Virol. 1994;137(3-4):389-95. doi: 10.1007/BF01309484.

Abstract

A recombinant baculovirus capable of expressing the glycoprotein H (gH) gene of equine herpesvirus 1 (EHV-1) was constructed. EHV-1 gH gene products in recombinant baculovirus infected insect cells were identified as 105 kDa and 110 kDa species compared with a 115 kDa product detected in EHV-1 infected mammalian cells. The extent of N-glycosylation of EHV-1 gH in both insect and mammalian cells was indicated by a shift in apparent molecular weights after PNGase F treatment to 90 kDa and 95 kDa forms, which compared with the predicted value of 90 kDa for the unglycosylated polypeptide. The recombinant EHV-1 gH was recognised by equine sera demonstrating that EHV-1 gH is a target for the immune system of the natural host. However, while the recombinant EHV-1 gH product from infected insect cells was immunogenic in mice, it did not induce a neutralizing antibody response against EHV-1.

摘要

构建了一种能够表达马疱疹病毒1型(EHV-1)糖蛋白H(gH)基因的重组杆状病毒。与在感染EHV-1的哺乳动物细胞中检测到的115 kDa产物相比,重组杆状病毒感染昆虫细胞中的EHV-1 gH基因产物被鉴定为105 kDa和110 kDa的种类。PNGase F处理后,昆虫和哺乳动物细胞中EHV-1 gH的N-糖基化程度通过表观分子量的变化得以体现,分别变为90 kDa和95 kDa的形式,与未糖基化多肽的预测值90 kDa相比。重组EHV-1 gH被马血清识别,表明EHV-1 gH是天然宿主免疫系统的靶标。然而,虽然来自感染昆虫细胞的重组EHV-1 gH产物在小鼠中具有免疫原性,但它并未诱导针对EHV-1的中和抗体反应。

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