Construction of a Series of C-terminal Truncation Mutants of Pseudorabies Virus VP22 and Their Subcellular Localization

YAO Lin; FANG Liu-rong*; XIAO Shao-bo; PAN Yong-fei; XIE Jing-guo; CHEN Huan-chun

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August 2006

YAO Lin, FANG Liu-rong*, XIAO Shao-bo, PAN Yong-fei, XIE Jing-guo and CHEN Huan-chun. Construction of a Series of C-terminal Truncation Mutants of Pseudorabies Virus VP22 and Their Subcellular Localization[J]. Virologica Sinica, 2006, 21(4): 358-363.

Citation: YAO Lin, FANG Liu-rong*, XIAO Shao-bo, PAN Yong-fei, XIE Jing-guo, CHEN Huan-chun. Construction of a Series of C-terminal Truncation Mutants of Pseudorabies Virus VP22 and Their Subcellular Localization .VIROLOGICA SINICA, 2006, 21(4) : 358-363.

伪狂犬病毒VP22蛋白C-端系列缺失突变体的构建及亚细胞定位分析

1.
华中农业大学动物医学院动物病毒室，武汉 430070

摘要

以绿荧光蛋白(GFP)为标记，构建了一系列伪狂犬病毒VP22蛋白的C-端缺失突变体与GFP融合表达的真核表达质粒，脂质体介导转染Hela细胞，通过荧光显微镜观察分析各个缺失突变体的亚细胞定位，发现伪狂犬病毒VP22蛋白与核定位有关的结构域在第60个到第90个氨基酸残基之间，第111个到第 159个氨基酸残基有可能与形成细胞核内的颗粒有关，与微管蛋白结合有关的结构域可能在第187 到第241个氨基酸残基之间。上述研究结果为进一步深入研究伪狂犬病毒VP22蛋白的结构与功能奠定了基础。
- 伪狂犬病毒
- , VP22
- , 缺失突变体
- , 亚细胞定位

Construction of a Series of C-terminal Truncation Mutants of Pseudorabies Virus VP22 and Their Subcellular Localization

1.
Laboratory of Animal Virology，College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China

Abstract

To study the specific subcellular localization of different regions of Pseudorabies virus (PrV) VP22, a series of mutants with the C-terminal truncations were amplified from a plasmid containing the full-length VP22 gene of the Ea strain. The resulting truncation constructs were fused to ORF encoding the green fluorescent protein (GFP) to generate eukaryotic expression plasmids expressing VP22 mutants and GFP fusion proteins. HeLa cells were transiently transfected with these fusion expression plasmids. By detecting the GFP fluorescence localization, several potential regions of PrV VP22 for specific subcellular localization were determined as follows: 60-90 amino acids (aa) of VP22 are required for nuclear targeting; 111-159 aa may be involved with the formation of particles in the nucleus; 187-241 aa are needed for binding to microtubules. These results lay foundation for further study on the structure and function of PrV VP22.
- Pseudorabies virus
- , VP22
- , Deletion mutants
- , Subcellular localization

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction of a Series of C-terminal Truncation Mutants of Pseudorabies Virus VP22 and Their Subcellular Localization

1. Laboratory of Animal Virology，College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China

Abstract: To study the specific subcellular localization of different regions of Pseudorabies virus (PrV) VP22, a series of mutants with the C-terminal truncations were amplified from a plasmid containing the full-length VP22 gene of the Ea strain. The resulting truncation constructs were fused to ORF encoding the green fluorescent protein (GFP) to generate eukaryotic expression plasmids expressing VP22 mutants and GFP fusion proteins. HeLa cells were transiently transfected with these fusion expression plasmids. By detecting the GFP fluorescence localization, several potential regions of PrV VP22 for specific subcellular localization were determined as follows: 60-90 amino acids (aa) of VP22 are required for nuclear targeting; 111-159 aa may be involved with the formation of particles in the nucleus; 187-241 aa are needed for binding to microtubules. These results lay foundation for further study on the structure and function of PrV VP22.