狂犬病病毒糖蛋白转基因小鼠的构建研究  

Study on Construction of Transgenic Mice Carrying Ra-bies Virus Glycoprotein cDNA

作　　者：扈荣良[1] 涂长春[1] 李红卫[1] 金扩世[1] 章金钢[1] 侯世宽[1] 崔青山[1] 殷震[1] 

机构地区：[1]长春农牧大学军事兽医研究所

出　　处：《中国兽医学报》1995年第3期209-213,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金;军队八五协作攻关课题

摘　　要：将狂犬病病毒糖蛋白ｃＤＶＡＢｇｌⅡ（１．６７ｋｂ）片段正向插入ｐＭＴ０１０／Ａ＋ＢａｍＨⅠ切点，构建重组质粒ｐＭＴ０１０／Ａ＋－Ｒｇｐ，用ＥｃｏＲⅠ＋ＳａｌⅠ对ｐＭＴ０１０／Ａ＋－Ｒｇｐ进行双酶切后，回收含有狂犬病病毒糖蛋白ｃＤＮＡ和绵羊ＭＴ启动子的２．７６ｋｂ片段，通过显微注射技术将该片段注入小鼠单细胞受精卵雄前核内，在进行胚胎移植后，获得４４只小鼠，经ＰＣＲ、Ｓｏｕｔｈ－ｅｒｎ杂交及原位杂交检测，证明有９只携带有目的基因。目的基因（即转基因）按孟德尔特性遗传给其后代，由此相应建立了９个转基因鼠系，对其中６个鼠系分别命名为ＴｇＮ（ｏＭＴ－Ｒｇｐ）ＩＬｇｅ、ＴｇＮ（ｏＭＴ－Ｒｇｐ）２Ｌｇｅ……ＴｇＮ（ｏＭＴ－Ｒｇｐ）６Ｌｇｅ。Expressing vector, pMT010/A+ -Rgp, was constructed by cloning rabies virus glycoprotein( Rgp)cDNA into downstream of the metallothionein promoter of pMT010/A+(ovine).The 2.76 kb Eco R I +Sal I fragments containing Rgp cDNA of pMT010/A+-Rgp were recovered and purified for microinjecting into male pronuclei of fertilized mouse eggs.Of 44 mice developed from these eggs, nine carried the gene of in- terest,which was identified by PCR, Southern hybridization and in situ hybridization. All of transgenic mice transmit their transgene to 50%of their offsprings indicative of Mendelian inheritance, Six out of 9 lines were designated as TgN (oMT- Rgp)1 Lge,TgN(oMT-Rgp)2Lge…TgN(oMT-Rgp)6Lge.

关 键 词：狂犬病 病毒 糖蛋白 转基因 小鼠 

分 类 号：S852.655[农业科学—基础兽医学]