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| 重组辛德毕斯病毒E基因痘苗病毒的构建 | |
| 引用本文： | 刘振江,刘昊,刘燕瑜,郭欢欢,凡敏,岳云强,陆飞,秦艳青,李国江,鲁会军,金宁一.重组辛德毕斯病毒E基因痘苗病毒的构建[J].中国兽医寄生虫病,2011,19(6):25-30. |
| 作者姓名： | 刘振江 刘昊 刘燕瑜 郭欢欢 凡敏 岳云强 陆飞 秦艳青 李国江 鲁会军 金宁一 |
| 作者单位： | 1. 中国人民解放军军事医学科学院军事兽医研究所,长春130122 吉林农业大学动物科技学院,长春130118 2. 中国人民解放军军事医学科学院军事兽医研究所,长春130122 吉林大学畜牧兽医学院,长春130062 3. 吉林农业大学动物科技学院,长春130118 吉林农业科技学院,吉林132101 4. 中国人民解放军军事医学科学院军事兽医研究所,长春,130122 |
| 摘 要： | 为了构建表达辛德毕斯病毒E基因的重组痘苗病毒,本研究通过基因重组的方法将辛德毕斯病毒E基因及绿色荧光蛋白（enhanced green fluorescent protein,EGFP）基因分别连接至痘苗病毒转移载体pSTK,酶切鉴定得到阳性重组质粒pSTK-SINE-EGFP。采用脂质体转染的方法,将该重组质粒与痘苗病毒天坛株共转染BHK-21细胞,通过同源重组获得重组痘苗病毒。利用EGFP筛选阳性重组痘苗病毒vTTVV-SINE-EGFP,收集感染重组痘苗病毒的BHK-21细胞,SDS-PAGE电泳检测辛德毕斯病毒E基因在细胞中的表达情况,Western blot分析表达产物的免疫原性。结果证明辛德毕斯病毒E基因能在重组痘苗病毒vTTVV-SINE-EGFP中获得表达,且表达产物具有良好的免疫原性。表达辛德毕斯病毒E基因的重组痘苗病毒的成功构建,为研制辛德毕斯病毒活载体疫苗奠定了基础。 |
| 关 键 词： | 辛德毕斯病毒 转移载体 绿色荧光蛋白 重组痘苗病毒 |
CONSTRUCTION OF RECOMBINANT VACCINIA VIRUS EXPRESSING E GENE OF SINDBIS VIRUS |
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| LIU Zhen-jiang,LIU Hao,LIU Yan-yuGUO Huan-huan,FAN Min,YUE Yun-qiang,LU Fei,QIN Yan-qing,LI Guo-jiang,LU Hui-jun,JIN Ning-yi.CONSTRUCTION OF RECOMBINANT VACCINIA VIRUS EXPRESSING E GENE OF SINDBIS VIRUS[J].Chinese Journal of Veterinary Parasitology,2011,19(6):25-30. | |
| Authors: | LIU Zhen-jiang LIU Hao LIU Yan-yuGUO Huan-huan FAN Min YUE Yun-qiang LU Fei QIN Yan-qing LI Guo-jiang LU Hui-jun JIN Ning-yi |
| Institution: | 1. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130122, China,2. College of Animal Science Jilin Agricultural University, Changchun 130118, China, 3. College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China ; 4. Jilin Agricaltural and Technology College, Jilin 132101, China) |
| Abstract: | To construct recombinant vaccinia virus expressing E gene of Sindbis virus, Sindbis virus E gene and green fluorescent protein （EGFP） gene were inserted to the vaccinia virus transfer vector pSTK by gene recombination. The resulting recombinant plasmid pSTK-SINE-EGFP was confirmed in restriction enzyme digestion. Subsequently, the recombinant plasmid and vaccinia virus Tiantan strain were co-transfected into BHK-21 cells and positive recombinant vaccinia virus was identified by fluorescent plaque purification. Sindbis virus E gene was expressed in BHK-21 cells infected with the recombinant vaccinia virus as determined in SDS-PAGE and Western blot. The results demonstrated that recombinant vaccinia virus expressing Sindbis virus E gene was constructed and expressed product showed good immunogenicity, which laid a solid foundation for development of a live vector vaccine of Sindbis virus. |
| Keywords: | Sindbis virus transfer vector enhanced green fluorescent protein （EGFP） recombinant vaccinia virus |
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