Construction and Identification of a Recombinant Canine Adenovirus Expressing Nucleocapsid Protein of Canine Distemper Virus

JU Hui-yan; XIA Xian-zhu; GAO Yu-wei; YANG Song-tao; ZOU Xiao-huan; HUANG Geng; LI Yan-fang

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August 2006

JU Hui-yan, XIA Xian-zhu, GAO Yu-wei, YANG Song-tao, ZOU Xiao-huan, HUANG Geng and LI Yan-fang. Construction and Identification of a Recombinant Canine Adenovirus Expressing Nucleocapsid Protein of Canine Distemper Virus[J]. Virologica Sinica, 2006, 21(4): 380-384.

Citation: JU Hui-yan, XIA Xian-zhu, GAO Yu-wei, YANG Song-tao, ZOU Xiao-huan, HUANG Geng, LI Yan-fang. Construction and Identification of a Recombinant Canine Adenovirus Expressing Nucleocapsid Protein of Canine Distemper Virus .VIROLOGICA SINICA, 2006, 21(4) : 380-384.

犬瘟热病毒核蛋白基因重组腺病毒的构建、鉴定与表达

鞠会艳 ^1,2 ,
夏咸柱 ^1*,, ,
高玉伟 ¹ ,
杨松涛 ¹ ,
邹啸环 ¹ ,
黄耕 ¹ ,
李彦舫 ²

1.
1.军事医学科学院军事兽医研究所，吉林长春 130062
2.
吉林大学农学部，吉林长春 130062

通讯作者： 夏咸柱, ;

摘要

核蛋白基因（N）位于犬瘟热病毒基因组的108—1679位置处，是保守性较强的免疫原性蛋白，因此选择N基因作为目的基因，利用酶切、连接等方法构建了含犬瘟热病毒核蛋白基因的穿梭质粒pVAXΔE3LPN。以含CAV-2 SY株全基因组的pPoly2-CAV-2为载体，构建了重组质粒pCAV-2-CDVLPN，利用脂质体介导法转染MDCK细胞，转染三次后，细胞出现了典型的腺病毒样病变。电镜负染、切片观察，酶切、PCR扩增及测序鉴定的结果表明表达犬瘟热核蛋白基因的重组犬2型腺病毒构建成功，表达的核蛋白分子量为58kDa。
- 犬瘟热病毒
- , 核蛋白
- , 犬腺病毒
- , 重组病毒

Construction and Identification of a Recombinant Canine Adenovirus Expressing Nucleocapsid Protein of Canine Distemper Virus

1.
1.Institute of Military Veterinary, Academy of Military Medical Science of PLA, Changchun 130062, China
2.
Department of Agricultural Science, Jilin University, Changchun 130062, China

Corresponding author: XIA Xian-zhu,

Abstract

The nucleocapsid protein gene (N) located 108 to 1679 situation of Canine distemper virus (CDV) genome and was immunogenicity protein with strong conservatism, so N gene was choosed as aim gene. A shuttle plasmid pVAXΔE3LPN containing the gene encoding the nucleocapsid protein of CDV was constructed by restriction endonuclease digestion and ligation. A recombinant plasmid pCAV-2-CDVLPN was constructed with pPoly2-CAV-2 vector containing CAV-2 SY genome. The plasmid pCAV-2-CDVLPN was transfected into MDCK cells by Lipofectamine and after three transfections, a typical CAV-2 cytopathic effect (CPE) was observed. We demonstrated that a recombinant canine adenovirus type-2 expressing nucleocapsid protein of canine distemper virus was successfully constructed and observed by electron microscopy, enzyme digestion, PCR and sequencing. The molecular mass of nucleocapsid protein was 58 kDa.
- Canine distemper virus (CDV)
- , Nucleocapsid Protein
- , Canine adenovirus (CAV)
- , Recombinant virus

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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 Identification of a Recombinant Canine Adenovirus Expressing Nucleocapsid Protein of Canine Distemper Virus

Corresponding author: XIA Xian-zhu,

1. 1.Institute of Military Veterinary, Academy of Military Medical Science of PLA, Changchun 130062, China
2. Department of Agricultural Science, Jilin University, Changchun 130062, China

Abstract: The nucleocapsid protein gene (N) located 108 to 1679 situation of Canine distemper virus (CDV) genome and was immunogenicity protein with strong conservatism, so N gene was choosed as aim gene. A shuttle plasmid pVAXΔE3LPN containing the gene encoding the nucleocapsid protein of CDV was constructed by restriction endonuclease digestion and ligation. A recombinant plasmid pCAV-2-CDVLPN was constructed with pPoly2-CAV-2 vector containing CAV-2 SY genome. The plasmid pCAV-2-CDVLPN was transfected into MDCK cells by Lipofectamine and after three transfections, a typical CAV-2 cytopathic effect (CPE) was observed. We demonstrated that a recombinant canine adenovirus type-2 expressing nucleocapsid protein of canine distemper virus was successfully constructed and observed by electron microscopy, enzyme digestion, PCR and sequencing. The molecular mass of nucleocapsid protein was 58 kDa.