[1]梅梅,陆吉虎,张雪花,等.高致病性血清4型禽腺病毒hexon基因在昆虫细胞中的表达及鉴定[J].江苏农业学报,2018,(04):862-865.[doi:doi:10.3969/j.issn.1000-4440.2018.04.021]
　MEI Mei,LU Ji-hu,ZHANG Xue-hua,et al.Expression and identification of high pathogenic fowl adenovirus serotype 4 hexon gene in insect cell[J].,2018,(04):862-865.[doi:doi:10.3969/j.issn.1000-4440.2018.04.021]
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高致病性血清4型禽腺病毒hexon基因在昆虫细胞中的表达及鉴定()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年04期
页码:
862-865
栏目:
畜牧兽医·水产养殖
出版日期:
2018-08-25

文章信息/Info

Title:
Expression and identification of high pathogenic fowl adenovirus serotype 4 hexon gene in insect cell
作者:
梅梅1; 2; 3; 陆吉虎1; 2; 3; 张雪花1; 2; 3; 唐应华1; 2; 3; 侯继波1; 2; 3
（1.江苏省农业科学院动物免疫工程研究所，江苏南京210014；2.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；3.江苏省动物重要疫病与人兽共患病防控协同创新中心，江苏扬州225009）
Author(s):
MEI Mei1; 2; 3; LU Ji-hu1; 2; 3; ZHANG Xue-hua1; 2; 3; TANG Ying-hua1; 2; 3; HOU Ji-bo1; 2; 3
（1.Institute of Veterinary Immunology and Engineering， Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.National Research Center of Engineering and Technology for Veterinary Biologicals, Jiangsu Academy of Agricultural Sciences, Nanjing 2
关键词:
高致病性血清4型禽腺病毒; 六邻体蛋白; hexon基因
Keywords:
high pathogenic fowl adenovirus serotype 4（FAdV-4）; hexon protein; hexon gene
分类号:
S855.3
DOI:
doi:10.3969/j.issn.1000-4440.2018.04.021
文献标志码:
A
摘要:
为获得杆状病毒表达的重组高致病性血清4型禽腺病毒（Fowl adenovirus serotype 4，FAdV-4）六邻体蛋白（Hexon），本研究通过PCR从高致病性血清4型禽腺病毒基因组中扩增到hexon基因，克隆到杆状病毒转移载体pFastBac-1，获得重组转移质粒pFastBac-hexon，将其转化DH10Bac大肠杆菌并筛选,获得重组杆状病毒穿梭质粒rBacmid-hexon,将重组杆状病毒穿梭质粒转染Sf9细胞，获得重组杆状病毒rBV-hexon。间接免疫荧光试验和免疫印迹试验结果显示，Sf9细胞中的Hexon蛋白能够与血清4型禽腺病毒阳性血清发生特异性反应，蛋白质分子量约110 000，表明Hexon蛋白在Sf9细胞中获得良好的表达。
Abstract:
To abtain the expression product of high pathogenic fowl adenovirus serotype 4（FAdV-4） hexon protein, hexon gene was amplified from genome of high pathogenic fowl adenovirus serotype 4, and it was cloned into transfer vector pFastBac-1. The recombinant transfer plasmid pFastBac-hexon was generated, transformed into DH10Bac Escherichia coli and screened to obtain recombinant shuttle plasmid rBacmid-hexon. The rBacmid-hexon was transfected into Sf9 cell to produce recombinant virus rBV-hexon. The expression of hexon was identified by indirect immunofluorescence assay and western blot, the results showed the recombinant virus in Sf9 cell can be recognized specifically by polyclonal antibody against FAdV-4, and the molecular weight was about 110 000, these results indicated that the hexon protein was well expressed in Sf9 cell.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期：2018-01-08 作者简介：梅梅（1982-），女，江苏东台人，博士，副研究员，主要从事分子病毒学和免疫学研究。（E-mail）jacqui18@163.com
更新日期/Last Update: 2018-09-04