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当前位置: 首页 > 文章 > 表达犬瘟热病毒核蛋白的重组犬2型腺病毒构建与包装 吉林农业大学学报 2005,27 (5)
Position: Home > Articles > Construction and Packaging of Recombinant Canine Adenovirus Type-2 Expression Nucleocapsid Protein of Canine Distemper Virus Journal of Jilin Agricultural University 2005,27 (5)

表达犬瘟热病毒核蛋白的重组犬2型腺病毒构建与包装

作  者：
鞠会艳;李彦舫;夏咸柱;高玉伟;杨松涛
单  位：
吉林大学农学部;军事医学科学院军事兽医研究所
关键词：
犬瘟热病毒;核蛋白(N蛋白);犬腺病毒;重组病毒
摘  要：
为构建表达犬瘟热病毒核蛋白的重组犬2型腺病毒,用MluⅠ和SphⅠ双酶切真核表达载体pVAXLPN,获得CDV N基因表达盒,将CDV N基因表达盒克隆入预先用SspⅠ酶切后的线性化质粒pVAXΔE3中,构建含CDV N基因表达盒的转移质粒pVAXΔE3LPN。用NruⅠ和SalⅠ双酶切转移质粒pVAXΔE3LPN,回收E3区部分缺失的含CDV N基因表达盒的目的片段,定向克隆入同样酶切处理含CAV-2全基因组的质粒载体ppoly2-CAV-2中,构建E3区部分缺失的包含CDV N基因表达盒的重组质粒pCAV-2-CDVLPN。重组质粒pCAV-2-CDVLPN经AscⅠ酶切获得线性化基因组CAV-2-CDVLPN。此基因组与NruⅠ和SalⅠ分别酶切CAV-2 SY基因组获得的片段混合,利用脂质体介导共转染MDCK细胞,重复转染3次后,出现典型CAV-2病变,经电镜观察和酶切鉴定,证明表达犬瘟热核蛋白的重组犬2型腺病毒包装成功。
译  名：
Construction and Packaging of Recombinant Canine Adenovirus Type-2 Expression Nucleocapsid Protein of Canine Distemper Virus
作  者：
JU Hui-yan~(1,2),LI Yan-fang~2,XIA Xian-zhu~1,GAO Yu-wei~1,YANG Song-tao~2 (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)
关键词：
canine distemper virus;nucleocapsid protein(N protein);canine adenovirus;recombinant virus
摘  要：
The expression cassette of the canine distemper virus nucleocapsid(CDV N) was released from pVAXLPN plasmid by digestion with MluⅠand SphⅠin order to construct recombinant canine adenovirus type-2(CAV-2) expression nucleocapsid protein of canine distempter virus.Then the CDV N expression cassette was blunted and ligated into(SspⅠ) site of pVAXΔE3 plasmid so that transferring vector plasmid pVAXΔE3LPN was constructed.After pVAXΔE3LPN plasmid was digested with NruⅠ and(SalⅠ),the objective fragment containing the CDV N expression cassette and some of adenovirus E3 regions were extracted and cloned into ppoly2-CAV-2 plasmid which was digested with the same enzyme in order to generate recombinant plasmid pCAV-2-CDVLPN.The linearization genosome CAV-2-CDVLPN was obtained after recombinant plasmid pCAV-2-CDVLPN was digested with AscⅠ.The CAV-2-CDVLPN was mixed with the segments of CAV-2 SY genosome digested with SalⅠand NruⅠ.Then the mixture was co-transfected into MDCK cells by Lipofectamine.After three transfections,typical CAV-2 cytopathic effect(CPE) was observed by microscope.It was proved that recombinant canine adenovirus type-2 expression nucleocapsid protein of canine distemper virus was successfully packed by electr on microscope observation and enzyme digestion.

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