【作者】 鞠会艳； 夏咸柱； 高玉伟； 杨松涛； 李彦舫；

【Author】 Ju Hui - Yan XIA Xian - Zhul Gao Yu - Wei YANG Song - Tao Li Yan - Fang(Institute of Military Veterinary, Academy of Military Medical Science of PLA, Changchun 130062) (Department of Agricultural Science, Jinlin University, Changchun 130062)

【机构】 军事医学科学院军事兽医研究所； 吉林大学农学部；

【摘要】 为构建含犬瘟热病毒核蛋白基因的重组载体质粒,用Mlu I和Sph I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Ssp I酶切后的线性化质粒pVAX△E3中,构建含CDV N基因表达盒的转移质粒pVAX△E3LPN。用NruI和Sal I双酶切转移质粒pVAX△E3LPN,回收E3区部分缺失的含CDV N基因表达盒的目的片段,定向克隆人同样酶切处理含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,该重组质粒经酶切鉴定正确。本研究为进一步研制含犬瘟热病毒核蛋白基因的重组病毒和研制有效预防犬瘟热暴发的新型重组疫苗奠定了基础。更多还原

【Abstract】 The CDV N expression cassette was released from pVAXLPN plasmid which was digested with Mlu I and Sph I in order to construct recombinant plasmid expression nucleocapsid protein of canine distemper virus . Then the CDV N expression cassette was blunted and ligated into SspI site of pVAX△E3 plasmid so that transferring vector plasmid pVAX△E3LPN was constructed . After pVAX△E3LPN plasmid was digested with Nru I and Sal I , the objective fragment containing the CDV N expression cassette and some of adenovirus E3 region was extracted and cloned into ppoly2 - CAV -2 plasmid which was digested with same enzyme in order to generate recombinant plasmid pCAV - 2·CDVLPN. It was proved that the recombinant plasmid expression nucleocapsid protein of canine distemper virus was proper by enzyme digestion . The base on studying recombinant virus expression nucleocapsid protein of canine distemper vims new type recombinant vaccine in order to effectually prevent the canine distemper virus eruption was established in this experiment .更多还原