Construction and Immunogenicity of Recombinant Canine Adenovirus Type 2 of Feline Distemper Virus VP2

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December 2005

Construction and Immunogenicity of Recombinant Canine Adenovirus Type 2 of Feline Distemper Virus VP2[J]. Virologica Sinica, 2005, 20(6): 637-641.

Citation: . Construction and Immunogenicity of Recombinant Canine Adenovirus Type 2 of Feline Distemper Virus VP2 .VIROLOGICA SINICA, 2005, 20(6) : 637-641.

表达猫瘟热病毒VP2蛋白重组腺病毒的构建及其免疫原性研究

杨松涛 ^1,2 ,
夏咸柱 ^1* ,
乔军 ¹ ,
常爽 ² ,
谢之景 ¹ ,
鞠会艳 ² ,
邹啸环 ¹

1.
1. 吉林长春军事医学科学院军事兽医研究所 2.吉林大学农学部中国工程院

摘要

为构建能表达FPV VP2蛋白的重组犬2型腺病毒(CAV-2)载体。首先用PCR方法从FPV GT-2株细胞培养物中扩增出了VP2蛋白基因,将其克隆到真核表达质粒pVAX1中构建了含有FPV vp2基因的表达盒(CMV- VP2-PolyA),将该表达盒酶切后定向克隆到含有CAV-2 E3区的穿梭质粒pVAX△E3中,构建出pVAX △E3VP2。用Sal I+Nru I双酶切pVAX△E3VP2,回收含有目的基因表达盒部分,将其定向克隆入含有CAV-2 全基因组的骨架质粒pPoly2-CAV-2中,构建了重组质粒pCAV-2-FPV-VP2。Cla I+Asc I酶切pCAV-2-FPV- VP2释放出重组基因组,以此转染MDCK细胞,获得了重组病毒CAV-2-VP2。该重组病毒能使MDCK细胞产生腺病毒样细胞病变。Western blot检测证实,该重组病毒能表达具有免疫学活性的VP2蛋白。该重组病毒可以有效地诱导免疫猫产生抗FPV和CAV-2抗体。本实验表明该重组病毒有可能成为一个FPV的疫苗株。
- 猫瘟热病毒
- , 猫泛自细胞减少症病毒
- , VP2蛋白
- , 重组腺病毒
- , 免疫原性

Construction and Immunogenicity of Recombinant Canine Adenovirus Type 2 of Feline Distemper Virus VP2

^1,2 ,
^1* ,
¹ ,
² ,
¹ ,
² ,
¹

Abstract

In order to construct a recombinant canine adenovirus type 2 (CAV-2) of the VP2 of Feline distemper virus or Feline panleukopenia virus (FPV), the vp2 gene fragment of FPV GT-2 strain was amplified by PCR and cloned into pVAX1 vector. The complete VP2 expression cassette was subcloned into the shuttle vector pVAXΔE3 and then inserted into the pPoly2-CAV-2 backbone vector that contains the complete genome of CAV-2. Recombinant viral genome was linearized by ClaI/AscI and transfected into MDCK cell. The recombinant CAV-2-VP2 was a-chieved through 4 passages in MDCK, which showed typical cytopathogenic effect (CPE). The expressed FPV VP-2 was detected in the infected MDCK cells of the recombinant CAV-2-VP-2 by using FPV polyclonal antibodies. The specific antibodies of VP-2 and CAV-2 were induced in cats by the recombinant CAV-2-VP-2. The results indicated that the recombinant CAV-2-VP-2 may have the potential to be used as a FPV vaccine strain.
- Feline distemper virus
- , Feline panleukopenia virus
- , VP2
- , Recombinant canine
- , adenovirus type 2

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction and Immunogenicity of Recombinant Canine Adenovirus Type 2 of Feline Distemper Virus VP2

^1,2 ,
^1* ,
¹ ,
² ,
¹ ,
² ,
¹

Abstract: In order to construct a recombinant canine adenovirus type 2 (CAV-2) of the VP2 of Feline distemper virus or Feline panleukopenia virus (FPV), the vp2 gene fragment of FPV GT-2 strain was amplified by PCR and cloned into pVAX1 vector. The complete VP2 expression cassette was subcloned into the shuttle vector pVAXΔE3 and then inserted into the pPoly2-CAV-2 backbone vector that contains the complete genome of CAV-2. Recombinant viral genome was linearized by ClaI/AscI and transfected into MDCK cell. The recombinant CAV-2-VP2 was a-chieved through 4 passages in MDCK, which showed typical cytopathogenic effect (CPE). The expressed FPV VP-2 was detected in the infected MDCK cells of the recombinant CAV-2-VP-2 by using FPV polyclonal antibodies. The specific antibodies of VP-2 and CAV-2 were induced in cats by the recombinant CAV-2-VP-2. The results indicated that the recombinant CAV-2-VP-2 may have the potential to be used as a FPV vaccine strain.