细小病毒非结构蛋白转基因小鼠模型建立

PREPARATION OF PARVOVIRAL NONSTRUCTURE PROTEIN TRANSGENIC MICE

将小鼠乳腺癌病毒启动子控制的细小病毒非结构蛋白基因(长5.7kb)氯化铯超速离心,纯化透析后用显微注射法导入C57BL/SJL F_1小鼠受精卵雄核,植入假孕母鼠输卵管,得成活小鼠15只。抽取鼠尾DNA,对其中10只小鼠作PCR和southern blot鉴定,其中4只(40％)整合有目的基因。对首建者B_6()的8只子代小鼠鉴定,3只(37.5％)整合有目的基因。说明导入的目的基因能传代。

DNA containiag the genes of parvoviral nonstructure protein was microinjected into the male pronucleus of fcrtilizecl ovum (C57,BL/SJL F1), the injected eggs (225) were implanted into the failopian tube of 9 pseudopregnnnt foster mothers (BABL/C). After a normai gesta-tion cnsues (approximately 3 weeks), 4 out of 9 faster mothers delivercd 15 viable pups. DNA was cxtracted from tails of the 10 of 15 pups. PCR and Southern blot Were performed to con-firm the presence of the NS gene. 4 out of the 10 mice were showed the integration of the NS gene. The 8 offspring of founder B 6 ( (?) ) were also checked by PCR and Southern blot, 3 out of the 8 mice were found the integration of NS gene, the results suggested that the integrated NS gene are inheritable.