| 表达传染性支气管炎病毒S1基因重组鸡痘病毒的构建 |
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| 引用本文： | 田占成,孙永科,王云峰,智海东,刘胜旺,王玫,童光志.表达传染性支气管炎病毒S1基因重组鸡痘病毒的构建[J].中国预防兽医学报,2006,28(4):466-470. |
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| 作者姓名： | 田占成 孙永科 王云峰 智海东 刘胜旺 王玫 童光志 |
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| 作者单位： | 1. 中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001;内蒙古农业大学,动物科技学院,内蒙古,呼和浩特,010018
2. 中国农业科学院哈尔滨兽医研究所,兽医生物技术国家重点实验室,黑龙江,哈尔滨,150001 |
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| 基金项目： | 高比容电子铝箔的研究开发与应用项目 |
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| 摘 要： | 将鸡传染性支气管炎病毒S1基因插入到鸡痘病毒转移载体pSY681中，获得重组转移载体pSY681。将pSY681-IBVS1转染已感染亲本鸡痘病毒S-FPV-017株的鸡胚成纤维细胞，使其在鸡胚成纤维细胞内与鸡痘病毒基因组发生同源重组，产生表达鸡IBVS1蛋白的重组鸡痘病毒rFPV-IBVS1。在含有X-gal的营养琼脂培养基上进行蓝斑筛选且进一步纯化14代。S1基因的PCR检测表明，获得的含传染性支气管炎病毒S1基因的重组鸡痘病毒能够稳定遗传，间接免疫荧光和Western blot等试验证实该重组病毒在CEF内真实地表达了分子量约为90Ku的具有免疫学活性的IBV S1糖蛋白。
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| 关 键 词： | 传染性支气管炎病毒 重组鸡痘病毒 S1基因 |
| 文章编号： | 1008-0589(2006)04-0466-05 |
| 收稿时间： | 2005-04-13 |
| 修稿时间： | 2005年4月13日 |
Construction of recombinant fowlpox virus expressing S1 gene of infectious bronchitis virus |
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| TIAN Zhan-cheng,SUN Yong-ke,WANG Yun-feng,ZHI Hai-Dong,LIU Sheng-Wang,WANG Mei,TONG Guang-Zhi.Construction of recombinant fowlpox virus expressing S1 gene of infectious bronchitis virus[J].Chinese Journal of Preventive Veterinary Medicine,2006,28(4):466-470. |
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| Authors: | TIAN Zhan-cheng SUN Yong-ke WANG Yun-feng ZHI Hai-Dong LIU Sheng-Wang WANG Mei TONG Guang-Zhi |
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| Institution: | 1. National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences Harbin 150001 ,China; 2. Inner Mongolia University of Agriculture, Huhhot 010018,China |
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| Abstract: | The S1 glycoprotein of infectious bronchitis virus could induce virus neutralization antibodies and play an important role in humoral protective immunity.To construct a recombinant fowlpox virus expressing S1 gene of infectious bronchitis virus,firstly the recombinant transferring vector PSY681-IBVS1 was constructed by inserting the full length infectious bronchitis virus S1 gene into the fowlpox virus transferring vector PSY681.Then the wide type fowlpox virus S-FPV-017 pre-infected chicken embryo fibroblasts(CEF)was transfected with PSY681-IBVS1.The recombinant fowlpox virus(rFPV-IBVS1)was selected with blue plaques on the CEF overlaid with X-gal agar.After fourteen generations selection,the recombinant fowlpox virus was purified.To identify the recombinant virus,PCR,indirect immunofluorescence and Western blot assay were carried out.The results demonstrated that the S1 gene of infectious bronchitis virus expressed was well and stably in recombinant fowlpox virus(rFPV-IBVS1). |
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| Keywords: | infectious bronchitis virus recombinant fowlpox virus Sl gene |
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