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伪狂犬病病毒ul24基因表达蛋白的胞内定位研究
引用本文:苏鑫铭 徐亚林 于春梅 曹瑞兵 周斌 陈溥言. 伪狂犬病病毒ul24基因表达蛋白的胞内定位研究[J]. Virologica Sinica, 2006, 21(5): 463-467
作者姓名:苏鑫铭 徐亚林 于春梅 曹瑞兵 周斌 陈溥言
作者单位: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(苏鑫铭,于春梅,曹瑞兵,周斌,陈溥言),南京医科大学微生物与免疫学教研室 江苏南京210029(徐亚林)
摘    要:
根据GenBank已发表的PrVul24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WesternBlot验证重组质粒。ul24基因序列测定结果已提交GenBank,登录号DQ226544。Westernblot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。

关 键 词:伪狂犬病病毒  PCR  ul24  GFP
收稿时间:2006-03-10
修稿时间:2006-04-17

Intracellular Localization of Pseudorabies Virus UL24 Protein
SU Xin-ming, XU Ya-lin, YU Chun-mei, CAO Rui-bing, ZHOU Bin, CHEN Pu-yan. Intracellular Localization of Pseudorabies Virus UL24 Protein[J]. 中国病毒学(英文版), 2006, 21(5): 463-467
Authors:SU Xin-ming   XU Ya-lin   YU Chun-mei   CAO Rui-bing   ZHOU Bin   CHEN Pu-yan
Affiliation: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.
Keywords:Pseudorabies virus  PCR  ul24  GFP  
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