传染性法氏囊病病毒VP2基因在昆虫细胞中的表达

›› 2013, Vol. 40 ›› Issue (9): 65-69.

传染性法氏囊病病毒VP2基因在昆虫细胞中的表达

杨凤, 黎先伟, 孙敏华, 严福强, 韦庆兰, 向斌, 任涛

华南农业大学兽医学院, 农业部新兽药创制重点开放实验室, 广东广州 510000

收稿日期:2013-03-04 出版日期:2013-09-20 发布日期:2013-09-18
通讯作者: 任涛 E-mail:rentao@scau.edu.cn
作者简介:杨凤(1988-),女,贵州人,硕士生,研究方向:禽病。
基金资助:
省科技计划项目(2012A020800006)。

Expression of VP2 Gene of Infectious Bursal Disease Virus in Insect Cells

YANG Feng, LI Xian-wei, SUN Min-hua, YAN Fu-qiang, WEI Qing-lan, XIANG Bin, REN Tao

Key Laboratory of New Drug Discovery, Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China

Received:2013-03-04 Online:2013-09-20 Published:2013-09-18

摘要/Abstract

摘要： 本研究将鸡传染性法氏囊病病毒超强毒(very virulent infectious bursal disease virus,vvIBDV)JM-1/10株VP2基因克隆至pcDNA-3.1(+)启动子CMV之后,随后将CMV-VP2基因序列一同克隆入杆状病毒表达系统的质粒 pFastBac^TM Daul中构建了pFast-CMV-VP2。将pFast-CMV-VP2转化Escherichia coli DH10Bac感受态细胞,筛选出重组质粒Bacmid-CMV-VP2。用 Bacmid-CMV-VP2转染Sf9昆虫细胞,获得了重组杆状病毒vBac-CMV-VP2。将该重组杆状病毒转导BHK-21细胞,48~72 h后经间接免疫荧光试验(IFA)检测到VP2蛋白具有特异性荧光;样品经Western blotting分析,结果显示目的蛋白得到表达。结果表明,本试验制备的重组 vBac-CMV-VP2 既能在昆虫细胞中表达,也可在哺乳动物细胞中表达。

关键词: 传染性法氏囊病病毒; VP2基因; 昆虫表达载体

Abstract: The VP2 gene of very virulent infectious bursal disease virus (vvIBDV) JM-1/10 strain was placed behind the CMV promoter of pcDNA-3.1(+).The CMV-VP2 gene was cloned into the baculovirus expression system pFastBac^TM Dual to build pFast-CMV-VP2.Transformed it to Escherichia coli DH10Bac.Recombinant baculovirus expression plasmid Bacmid-CMV-VP2 was screened.Bacmid-CMV-VP2 was used to be transfected into Sf9 insect cells to get the recombinant baculovirus vBac-CMV-VP2 and transduced into BHK-21 cells.After 48 to 72 hours,indirect immunofluorescence assay (IFA) was conducted to detect specific fluorescence.Western blotting analysis result showed target protein was expressed.The prepared recombinant vBac-CMV-VP2 could not only be expressed in insect cells, but also expressed in mammalian cells.

Key words: infectious bursal disease virus; VP2 gene; insect expression vector

中图分类号:

Q786

杨凤, 黎先伟, 孙敏华, 严福强, 韦庆兰, 向斌, 任涛. 传染性法氏囊病病毒VP2基因在昆虫细胞中的表达[J]. , 2013, 40(9): 65-69.

YANG Feng, LI Xian-wei, SUN Min-hua, YAN Fu-qiang, WEI Qing-lan, XIANG Bin, REN Tao. Expression of VP2 Gene of Infectious Bursal Disease Virus in Insect Cells[J]. China Animal Husbandry & Veterinary Medicine, 2013, 40(9): 65-69.

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