利用转基因小鼠表达法氏囊病毒A片段的研究  

Study on Expressing A Segment of Infectious Bursal Deasease Virus to Milk in Transgenic Mice

作　　者：林爱星[1] 周云[1] 胡宏宇[1] 侯建[1] 安晓荣[1] 陈永福[1] 

机构地区：[1]中国农业大学农业生物技术国家重点实验室,北京100094

出　　处：《农业生物技术学报》2003年第3期280-284,共5页Journal of Agricultural Biotechnology

基　　金：国家"863"高技术计划基金。

摘　　要：将法氏囊病毒(IBDV)A片段定向连于乳腺特异性表达载体p7GMB53,用于转基因实验,用PCR和Southern杂交对阳性小鼠及其后代进行检测。共注射了879枚小鼠受精卵,移植受体26只,产仔39只。经PCR检测阳性6只(其中2只死胎),Southern杂交检测确定了2只阳性小鼠(1公1母)。外源基因整合的拷贝数分别为3拷贝(公)和9拷贝(母)。将两只阳性小鼠进行了传代,实验表明二者分别为种系纯合体及种系嵌合体。ELISA及Westernblot检测认为阳性母鼠表达了IBDV衣壳蛋白。初步验证了先前构建的含基质附着区序列(MARs)的乳腺表达载体能克服转基因的位置效应,实现转入目的基因的表达,提高乳腺反应器的制备效率。The A segment of infectious bursal desease virus (IBDV) was ligated to the mammary gland expression vector p7GMB53. This transgenic construct was linearized to perform transgenic experiments, total 879 fertilized eggs were microinjected and transferred into 26 recipient mice, 39 mice were born with 4.4%(39/879) embryo survival rate. Results of PCR showed that 4 of 39 were positive. Southern hybridization showed 2 of 4 were positive, in which the copy numbers of IBDV integrated were 3(No.8) and 9(No.3) respectively. The female mouse No.3 was mated with nontransgenic male mouse and transgenic male mouse No.8. The ratio of positive mice was 52% (10/19) and 100%(5/5) respectively and the result indicated that the No.3 was a germ pure line. The No.8 was mated with nontransgenic female mouse and the ratio of positive mice was 11%(2/18), indicated the No.8 was a germ chimera. The milks of No.3 (female), No.6(the female offspring of No.3 and a nontransgenic mouse) and No.10 (the female offspring of the No.3 and No.8) were performed by Western blot and ELISA. Both of the mice expressed the foreign gene in mammary glands. We concluded preliminarily that this vector, containing matrix attachment region seguence(MARs), could drive foreign gene to express to milk independent effects of position in transgenic animals.

关 键 词：位置效应 基质附着区序列 转基因小鼠 法氏囊病毒A片段 表达载体 鸡传染性法氏囊病毒 

分 类 号：S858.31[农业科学—临床兽医学] S852.65[农业科学—兽医学]