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pVAX?E3LPN。以含CAV-2SY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。
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 pVAXAE3LPN 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.
出处
《中国病毒学》
CAS
CSCD
2006年第4期380-384,共5页
Virologica Sinica
基金
全军医药卫生科研基金课题01-Z-092
关键词
犬瘟热病毒
核蛋白
犬腺病毒
重组病毒
Canine distemper virus (CDV)
Nucleocapsid Protein
Canine adenovirus (CAV)
Recombinant virus