[1]许梦微,王志胜,乔永峰,等.表达禽流感病毒H5亚型基因重组鸭瘟病毒的构建[J].江苏农业学报,2015,(06):1371-1377.[doi:doi:10.3969/j.issn.1000-4440.2015.06.027]
　XU Meng-wei,WANG Zhi-sheng,QIAO Yong-feng,et al.Construction of a vectored duck enteritis virus (DEV) expressing hemagglutinin of H5N1 avian influenza virus[J].,2015,(06):1371-1377.[doi:doi:10.3969/j.issn.1000-4440.2015.06.027]
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表达禽流感病毒H5亚型基因重组鸭瘟病毒的构建()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年06期
页码:
1371-1377
栏目:
畜牧兽医·水产养殖
出版日期:
2015-12-31

文章信息/Info

Title:
Construction of a vectored duck enteritis virus (DEV) expressing hemagglutinin of H5N1 avian influenza virus
作者:
许梦微12; 王志胜1;  乔永峰1; 顾一奇12 ; 柳畅12; 侯继波1 ; 姜平2; 王继春1
（1.江苏省农业科学院国家兽用生物制品工程技术研究中心，江苏南京210014；2.南京农业大学动物医学院，江苏南京210095）
Author(s):
XU Meng-wei12; WANG Zhi-sheng1; QIAO Yong-feng1; GU Yi-qi12; LIU Chang12; HOU Ji-bo1; JIANG Ping2; WANG Ji-chun1
(1.National Research Center of Veterinary Biologicals Engineering and Technology, Jiangsu Academy of Agricultural Sciences,Nanjing 210014, China；
关键词:
重组鸭瘟病毒; 细菌人工染色体; 高致病禽流感病毒; HA基因
Keywords:
recombination duck enteritis virus; bacterial artificial chromosome; avian influenza virus; HA gene
分类号:
S858.32
DOI:
doi:10.3969/j.issn.1000-4440.2015.06.027
文献标志码:
A
摘要:
为了应用和发展鸭肠炎病毒（DEV）载体疫苗，基于细菌人工染色体（BAC）人工合成带有pMCMV IE启动子和HA基因序列的表达盒，将其插入到UL55和LORF11之间的非编码区，从而构建出DEV载体疫苗。RFLP分析结果显示DEV BAC及其突变体条带与预期结果存在轻微的差异，将BAC及其突变体拯救后发现其生长性能与亲本病毒没有显著差异。间接免疫荧光和Western blot结果显示，载体疫苗在感染鸡胚成纤维细胞（CEF）后大量表达了HA蛋白质。 表明成功构建了能稳定表达禽流感病毒H5亚型基因重组鸭瘟病毒。
Abstract:
To facilitate the generation and further improvement of duck enteritis virus(DEV) vectored HA(H5) vaccine, based on the bacterial artificial chromosome (BAC), a synthesized HA(H5) expression cassette with pMCMV IE promoter and a consensus HA sequences were inserted into the non-coding area between the UL55 and LORF11. Restriction fragment length polymorphism (RFLP) of DEV BAC and its mutants showed slight differences with expected result. Recovered virus from the BAC or mutants exhibited similar growth kinetics as parental virus. The results of indirect immunofluorescence and Western blotting revealed the robust expression of HA infected by the vectored vaccine in chicken embryo fibroblasts(CEFs). It was indicated that a vectored duck enteritis virus (DEV) stably expressing hemagglutinin of H5N1 avian influenza virus were successfully constructed based on an infectious clone of DEV vaccine strain.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2015-03-17 基金项目：公益性行业(农业)科研专项 (201303046)； 江苏省农业科技自主创新基金项目[CX(14)2048];江苏省自然科学基金项目(BK20131334) 作者简介：许梦微（1988-），女，江西抚州人，硕士研究生，主要从事病毒载体活疫苗的研究。(Tel)18151689053；(E-mail)xumengwei1746109@aliyun.com 通讯作者：王继春，(E-mail)jcwang@263.net
更新日期/Last Update: 2015-12-31