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利用基于马铃薯Y病毒载体在植物中表达的VP60蛋白免疫家兔,使其免受兔出血症病毒感染。

Protection of rabbits against rabbit hemorrhagic disease virus by immunization with the VP60 protein expressed in plants with a potyvirus-based vector.

作者信息

Fernández-Fernández M R, Mouriño M, Rivera J, Rodríguez F, Plana-Durán J, García J A

机构信息

Centro Nacional de Biotecnología-CSIC, Campus de la Universidad Autónoma de Madrid, 28049 Madrid, Spain.

出版信息

Virology. 2001 Feb 15;280(2):283-91. doi: 10.1006/viro.2000.0762.

Abstract

A new plum pox potyvirus (PPV)-based vector has been constructed for the expression of full-length individual foreign proteins. The foreign sequences are cloned between the NIb replicase and capsid protein (CP) cistrons. The heterologous protein is split from the rest of the potyviral polyprotein by cleavage at the site that originally separated the NIb and CP proteins and at an additional NIa protease recognition site engineered at its amino-terminal end. This vector (PPV-NK) has been used to clone different genes, engendering stable chimeras with practical applications. We have constructed a chimera expressing high levels of jellyfish green fluorescent protein, which can be very useful for the study of PPV molecular biology. The VP60 structural protein of rabbit hemorrhagic disease virus (RHDV) was also successfully expressed by making use of the PPV-NK vector. Inoculation of extracts from VP60-expressing plants induced a remarkable immune response against RHDV in rabbits, its natural host. Moreover, these animals were protected against a lethal challenge with RHDV.

摘要

一种基于李痘病毒(PPV)的新型载体已构建成功,用于全长单个外源蛋白的表达。外源序列克隆于NIb复制酶和顺反子衣壳蛋白(CP)之间。异源蛋白通过在最初分隔NIb和CP蛋白的位点以及在其氨基末端设计的额外NIa蛋白酶识别位点处进行切割,从马铃薯Y病毒多聚蛋白的其余部分中分离出来。该载体(PPV-NK)已用于克隆不同基因,产生具有实际应用价值的稳定嵌合体。我们构建了一种表达高水平水母绿色荧光蛋白的嵌合体,这对于PPV分子生物学的研究可能非常有用。利用PPV-NK载体还成功表达了兔出血性疾病病毒(RHDV)的VP60结构蛋白。接种表达VP60的植物提取物可在RHDV的天然宿主兔中诱导出显著的针对RHDV的免疫反应。此外,这些动物受到保护,可抵御RHDV的致死性攻击。

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