Intracellular Localization of Pseudorabies Virus UL24 Protein

SU Xin-ming; XU Ya-lin; YU Chun-mei; CAO Rui-bing; ZHOU Bin; CHEN Pu-yan

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October 2006

SU Xin-ming, XU Ya-lin, YU Chun-mei, CAO Rui-bing, ZHOU Bin and CHEN Pu-yan. Intracellular Localization of Pseudorabies Virus UL24 Protein[J]. Virologica Sinica, 2006, 21(5): 463-467.

Citation: SU Xin-ming, XU Ya-lin, YU Chun-mei, CAO Rui-bing, ZHOU Bin, CHEN Pu-yan. Intracellular Localization of Pseudorabies Virus UL24 Protein .VIROLOGICA SINICA, 2006, 21(5) : 463-467.

伪狂犬病病毒ul24基因表达蛋白胞内定位研究

苏鑫铭 ¹ ,
徐亚林 ² ,
于春梅 ¹ ,
曹瑞兵 ¹ ,
周斌 ¹ ,
陈溥言 ^1*

1.
1. 南京农业大学农业部动物疫病诊断与免疫重点开放实验室江苏南京 210095
2.
南京医科大学微生物与免疫学教研室江苏南京 210029

摘要

根据GenBank已发表的PrV ul24基因序列(NC006151)，设计并合成一对引物，PCR扩增出ul24基因编码区，克隆于pEGFP-N1载体，得到重组质粒pUL24-GFP。酶切鉴定，测序及Western Blot验证重组质粒。ul24基因序列测定结果已提交GenBank，登录号DQ226544。Western blot分析结果表明UL24-GFP融合蛋白为45KD。将pUL24-GFP转染真核细胞，激光共聚焦显微镜观察融合蛋白的细胞内定位，结果表明UL24-GFP融合蛋白定位于细胞核。
- 伪狂犬病病毒
- , PCR
- , ul24
- , GFP

Intracellular Localization of Pseudorabies Virus UL24 Protein

1.
1. Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
2.
Department of Microbiology and Immunology, Nanjing Medical University, Nanjing 210029, China

Abstract

A pair of primers were designed based on the PrV ul24 gene sequence in GenBank (NC006151). The UL24 protein coding sequence was amplified by PCR from PrV RongA strain genomic DNA. The product was cloned into pEGFP-N1 vector to generate the plasmid pUL24-GFP. Restriction endonuclease digestion, DNA sequencing and Western blots were employed to identify and authenticate pUL24-GFP. The ul24 DNA sequencing result was submitted to GenBank (DQ226544). Western blot analysis indicated that the UL24-GFP fusion protein was 45KD. After transfection of pUL24-GFP into eukaryotic cells, the intracellular localization of UL24-GFP fusion protein was examined by confocal microscopy and the result indicated that the fusion protein was localized mainly in nucleus.
- Pseudorabies virus
- , PCR
- , ul24
- , GFP

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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

Intracellular Localization of Pseudorabies Virus UL24 Protein

1. 1. Key Laboratory of Animal Diseases Diagnosis and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
2. Department of Microbiology and Immunology, Nanjing Medical University, Nanjing 210029, China

Abstract: A pair of primers were designed based on the PrV ul24 gene sequence in GenBank (NC006151). The UL24 protein coding sequence was amplified by PCR from PrV RongA strain genomic DNA. The product was cloned into pEGFP-N1 vector to generate the plasmid pUL24-GFP. Restriction endonuclease digestion, DNA sequencing and Western blots were employed to identify and authenticate pUL24-GFP. The ul24 DNA sequencing result was submitted to GenBank (DQ226544). Western blot analysis indicated that the UL24-GFP fusion protein was 45KD. After transfection of pUL24-GFP into eukaryotic cells, the intracellular localization of UL24-GFP fusion protein was examined by confocal microscopy and the result indicated that the fusion protein was localized mainly in nucleus.