[1]王安平,朱善元,王永娟,等.鸭源新城疫病毒NP基因在昆虫细胞中的表达及抗原性检测[J].江苏农业学报,2015,(06):1384-1388.[doi:doi:10.3969/j.issn.1000-4440.2015.06.029]
　WANG An-ping,ZHU Shan-yuan,WANG Yong-juan,et al.Expression and antigenicity of NP gene of duck Newcastle disease virus （NDV）in insect cells[J].,2015,(06):1384-1388.[doi:doi:10.3969/j.issn.1000-4440.2015.06.029]
点击复制

鸭源新城疫病毒NP基因在昆虫细胞中的表达及抗原性检测()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年06期
页码:
1384-1388
栏目:
畜牧兽医·水产养殖
出版日期:
2015-12-31

文章信息/Info

Title:
Expression and antigenicity of NP gene of duck Newcastle disease virus （NDV）in insect cells
作者:
王安平; 朱善元; 王永娟; 吴双; 左伟勇; 洪伟鸣
（江苏农牧科技职业学院江苏省兽用生物制药高技术研究重点实验室，江苏泰州225300）
Author(s):
WANG An-ping; ZHU Shan-yuan; WANG Yong-juan; WU Shuang; ZUO Wei-yong; HONG Wei-ming
(Jiangsu Provincial Key Laboratory of Veterinary Bio-pharmaceutical High-tech Research， Jiangsu Agri-animal Husbandry Vocational College,Taizhou 225300，China)
关键词:
鸭源新城疫病毒; NP基因; 昆虫细胞; 表达
Keywords:
duck Newcastle disease virus; NP gene; insect cell; expression
分类号:
S834
DOI:
doi:10.3969/j.issn.1000-4440.2015.06.029
文献标志码:
A
摘要:
为在昆虫细胞中表达鸭源新城疫病毒NP蛋白质，首先根据鸭源新城疫病毒NP基因序列设计引物，扩增出NP基因，将其克隆至杆状病毒表达载体pFastBac1，获得重组转移载体pFastBac-NP，将其转化到DH10Bac感受态细胞中，经抗性和蓝白斑筛选，获得重组穿梭质粒rBacmid-NP，在脂质体的介导下转染昆虫细胞Sf9，获得重组杆状病毒NP蛋白质。Western blot、间接免疫荧光试验结果均显示表达的重组蛋白质能够与全病毒阳性血清发生特异性反应，具有良好的反应原性。以上结果说明鸭源新城疫病毒NP蛋白质在昆虫细胞中获得了成功表达。
Abstract:
To express the NP gene of duck NDV （Newcastle disease virus） in insect cells, one pair of specific primers were designed for the amplification of NP gene by PCR, and the amplified fragment was cloned into baculovirus expression vector pFastBac1. The recombinant vector pFastBac-NP was transformed into DH10Bac Escherichia coli, and the positive recombinant bacmid rBacmid-NP was screened according to the resistance and the blue-white plague screening. The NP protein was aquired through the transfection of rBacmid-NP into Sf9 insect cells by liposome. The results of Western blot and indirect immunofluorescence showed that the recombinant proteins could be recognized by positive serum, indicating that the protein had good reactiongenicity. These results suggested that the NP proteins of duck NDV had been successfully expressed in insect cells.

参考文献/References:

[1]王永坤,田慧芳,周继宏,等. 鹅副粘病毒病的研究[J]. 中国家禽, 1998, 20(4): 3-5.
[2]李世江,赵忠民,刘红娟,等. 鸭副粘病毒病的诊治[J]. 中国动物保健, 2003, 53(7): 51.
[3]何永强,洪健,吴旧生,等. 致麻鸭产蛋下降的副粘病毒的分离和鉴定[J]. 中国兽医学报, 2005, 25(3): 238-240.
[4]张耀成,朱汉华. 鸭副粘病毒与鸭疫里默氏杆菌混合感染的诊治[J]. 禽病防制, 2005, 22(5): 28.
[5]于洋,封振,何孔旺,等. 新城疫病毒毒力分子机制研究进展[J]. 江苏农业学报,2013,29(2): 435-439.
[6]吴双,王永娟,左伟勇,等. 新城疫病毒 LaSota 疫苗株的准种遗传变异分析[J]. 江苏农业科学,2014,42(3): 161-163.
[7]KHO C L, TAN W, YUSOFF K. Production of the nucleocapsid protein of newcastle disease virus in Escherichia coli and its assembly of into ring-like and nucleocapsid-like particles[J]. Journal of Microbiology, 2001, 39(4): 293-299.
[8]MAKKAY A M, KRELL P J, NAGY E. Antibody detection-based differential elisa for NDV-infected or vaccinated chickens versus NDV HN-subunit vaccinated chickens[J]. Veterinary Microbiology, 1999, 66(3): 209-222.
[9]HU Y C. Baculovirus as a highly efficient expression vector in insect and mammalian cells[J]. Acta Pharmacol Sin, 2005, 26(4): 405-416.
[10]MARANGA L, CRUZ P E, AUNINS J G, et al. Production of core and virus-like particles with baculovirus infected insect cells[J]. Adv Biochem Eng Biotechno, 2002, 74: 183-206.
[11]ZHANG T, XU Y, QIAO L, et al. Trivalent human papillomavirus (HPV ) VLP vaccine covering HPV type 58 can elicit high level of humoral immunity but also induce immune interference among component types[J]. Vaccine, 2010, 28(19): 3479-3487.

备注/Memo

备注/Memo:
收稿日期：2015-03-27 基金项目：国家自然科学基金项目（31302096）；江苏省农业科技支撑计划项目（BE2013415）；江苏省“六大人才高峰”项目（NY-009） 作者简介：王安平（1980-），女，江苏泰州人，博士，副教授，主要从事兽用生物制药的研究。（Tel）15189910087；（E-mail）wap4017@163.com 通讯作者：朱善元，（Tel）13775665658
更新日期/Last Update: 2015-12-31