[1]郑金,陈滔,陆吉虎,等.减蛋下降综合征病毒重组Knob-S基因在杆状病毒系统中的表达及其免疫原性鉴定[J].江苏农业学报,2015,(03):600-603.[doi:10.3969/j.issn.1000-4440.2015.03.022]
　ZHENG Jin,CHEN Tao,LU Ji-hu,et al.Expression of Knob-S gene of egg drop syndrome virus in baculovirus system and its immunogenicity[J].,2015,(03):600-603.[doi:10.3969/j.issn.1000-4440.2015.03.022]
点击复制

减蛋下降综合征病毒重组Knob-S基因在杆状病毒系统中的表达及其免疫原性鉴定()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年03期
页码:
600-603
栏目:
畜牧兽医·水产养殖
出版日期:
2015-06-30

文章信息/Info

Title:
Expression of Knob-S gene of egg drop syndrome virus in baculovirus system and its immunogenicity
作者:
郑金1; 陈滔12; 陆吉虎2; 高峰12; 候继波2; 黎满香1; 唐应华2
(1.湖南农业大学动物医学院，湖南长沙410128；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)
Author(s):
ZHENG Jin1; CHEN Tao12; LU Ji-hu2; GAO Feng12; HOU Ji-bo2; LI Man-xiang1; TANG Ying-hua2
（1.College of Veterinary Medicine, Hunan Agricultural University, Changsha410128, China;2.Jiangsu Academy of Agricultural Sciences/National Research Center of Engineering and Technology for Veterinary Biologicals, Nanjing210014, China）
关键词:
减蛋下降综合征病毒; 重组Knob-S; 真核表达; 杆状病毒表达系统
Keywords:
egg drop syndrome virus; recombinant Knob-S; eukaryotic expression; baculovirus expression system
分类号:
Q786
DOI:
10.3969/j.issn.1000-4440.2015.03.022
文献标志码:
A
摘要:
为以杆状病毒系统在Sf9细胞中表达含减蛋下降综合征病毒（EDSV）重组Knob-S蛋白质来制备亚单位疫苗，采用PCR方法从鸭胚繁殖的EDSV尿囊液中扩增Knob-S基因，并克隆到pFastBac1中，经转化DH10Bac获得重组rBac-Knob-S。将rBac-Knob-S转染Sf9细胞获得重组杆状病毒，利用重组杆状病毒感染Sf9细胞表达重组蛋白质。重组Knob-S蛋白质经Western blotting和间接免疫荧光试验鉴定均为阳性。将重组Knob-S蛋白质制备成疫苗，按1羽0.3 ml（含50 μg的重组病毒）免疫6周龄非免海兰褐雏鸡，在免疫后第2周可检测到血凝抑制抗体效价达8.5 lg2，在第3周达到高峰（9.5 lg2），表明重组Knob-S蛋白质具有较好的免疫原性。
Abstract:
To develop egg drop syndrome virus subunit vaccine (EDSV) based on its recombinant Knob-S proteins expressed in Sf9 cell, the Knob-S gene of EDSV was amplified from allantoic fluids of duck embryos by PCR, and cloned into pFastBac-1 before transformation into DH10Bac to construct rBac-Knob-S bacmid. The recombinant Knob-S proteins were expressed in Sf9 cell by infection of the recombinant baculovirus derived from the transfection of the bacmids in Sf9 cell. The recombinant Knob-S proteins were identified by Western blotting and indirect immunofluorescence assay. Groups of 6-week-old nonimmuned HY-Line brown chickens were inoculated with vaccines containing the recombinant Knob-S proteins. The serum antibody levels reached 8.5 lg2 2 week post-vaccination, and rose to the peak of 9.5 lg2 3 week post-vaccination determined by hemagglutinin inhibition test, indicating that recombinant Knob-S proteins had good immunogenicity for chickens. 

参考文献/References:


[1]W 卡尼尔 B. 禽病学[M]. 10版. 北京: 中国农业大学出版社, 1999.
[2]TOMKO R P，JOHANSSON C B，TOTROV M，et al. Expression of the adenovirus receptor and its interaction with the fiber knob[J]. Experimental Cell Research，2000，255(1)：47-55.
[3]FINGERUT E，GUTTER B，GALLILI G，et al. A subunit vaccine against the adenovirus egg-drop syndrome using part of its fiber protein[J]. Vaccine，2003，21(21)：2761-2766.
[4]陈滔，唐应华，陆吉虎，等. 产蛋下降综合征病毒纤突蛋白 Knob-S 区原核表达及其活性分析[J]. 动物医学进展，2012，33(6)：41-44.
[5]KOST T A，CONDREAY J P，JARVIS D L. Baculovirus as versatile vectors for protein expression in insect and mammalian cells[J]. Nature Biotechnology，2005，23(5)：567-575.
[6]SHEN X，HU G，JIANG S，et al. Engineering and characterization of a baculovirus-expressed mouse/human chimeric antibody against transferrin receptor[J]. Protein Engineering Design and Selection，2009，22(12)：723-731.
[7]WANG Y，SUN Y，TIAN Z，et al. Protection of chickens against infectious bronchitis by a recombinant fowlpox virus co-expressing IBV-S1 and chicken IFNγ[J]. Vaccine，2009，27(50)：7046-7052.
[8]NAGATA T，ISHIKAWA S，SHIMOKAWA E，et al. High level expression and purification of bioactive bovine interleukin-18 using a baculovirus system[J]. Veterinary Immunology and Immunopathology，2002，87(1)：65-72.
[9]唐应华，吴培培，宫玉珍,等. 肾型传染性支气管炎病毒的鉴定和结构蛋白的表达[J]. 中国兽医科学，2011，41(10)：1034-1040.
[10]李东卫，郭莹莹，刘在斯,等. 禽流感病毒 NS1 蛋白在杆状病毒表达系统中的表达[J]. 动物医学进展，2012，33(6)：27-31.〖ZK）〗〖FL）〗

备注/Memo

备注/Memo:
收稿日期：2015-01-13 基金项目：国家自然科学基金项目 (31201904)；农业部公益性行业（农业）科研专项经费项目（201303046）；科技部国家高技术研究发展计划“863”课题（2011AA10A213）；江苏省农业科技自主创新基金项目[CX(14)5045] 作者简介：郑金（1989-），女，湖南长沙人，硕士研究生，主要从事病原与分子生物学研究。（Tel）18390990241;（E-mail）296921762@qq.com 通讯作者：黎满香，（E-mail）manxiangl@163.com;唐应华，（E-mail）tyinghua@yhoo.com
更新日期/Last Update: 2015-06-30