鸡减蛋综合征病毒分离株AA－2基因组Hind　Ⅲ酶切部分片段的分子克隆  

Molecular Cloning of Hind Ⅲ Fragments of Egg Drop Syndrome Virus Genome

作　　者：胡敬东[1] 章金钢[1] 陶全福 李红卫[1] 扈荣良[1] 金扩世[1] 涂长春[1] 殷震[1] 

机构地区：[1]解放军农牧大学军事兽医研究所

出　　处：《中国兽医学报》1996年第3期244-247,共4页Chinese Journal of Veterinary Science

摘　　要：以非免疫鸭胚增殖的减蛋综合征病毒（ＥＤＳＶ），经差速离心法纯化，电镜下观察到病毒无囊膜，大小为７０～８５ｎｍ。以蛋白酶Ｋ法抽提病毒基因组，琼脂糖凝胶电泳只出现１条分子量约３４ｋｂ、背景清晰的核酸带。ＨｉｎｄⅢ酶切ＥＤＳＶＤＮＡ可见１０条片段，以ｐＵＣ１９质粒为载体，采用鸟枪法对病毒基因组的ＨｉｎｄⅢ酶切片段进行分子克隆，获得１７个重组质粒，通过酶切、斑点杂交鉴定，证实已经成功地克隆到ＥＤＳＶ的５．２４ｋｂ、２．０２ｋｂ、１．６５ｋｂ、１．４７ｋｂ、１．４１ｋｂ等５个ＤＮＡ片段。By using different rate centrifugations, egg drop syndrome viruses(EDSV) were purified from allantoic fluids of duck embryos inoculated with the EDSVstrain AA-2. Under electron microscope,the round virus particles were without envelopand calculated to be 70～85 nm in size.Viral DNA was extracted and purified,a clearband of about 34 kb in the agarose gel electrophoresis was found.The viral genomicDNA digested with Hind Ⅲ and showed 10 fragments.EDSV DNA Hind Ⅲ fragmentswere inserted into the Hind Ⅲ site of pUC19 by shot gun method and 17 recombinantplasmids were obtained. The recombinants were identified by digestion with Hind Ⅲand dot blot hybridization and 5 fragments of EDSV DNA in sizes of 5.24 kb,2.02 kb,1.65 kb,1.47 kb,1.41 kb respectively had been successfully cloned.The physicalmaps of the cloned framents are being studied.

关 键 词：EDSV 克隆 斑点杂交 

分 类 号：Q78[生物学—分子生物学] S852.65[农业科学—基础兽医学]