人Bcl-2乳腺定位表达转基因小鼠的建立  

Establishment of Transgenic Mice Expressing Bcl-2 in Mammary Gland

作　　者：赵君[1] 任文陟[1] 张守峰[1] 刘彬[2] 宇丽[3] 扈荣良[1] 殷震[1] 

机构地区：[1]解放军军需大学军事兽医研究所解放军基因工程重点实验室,吉林长春130062 [2]湖南大学,湖南长沙410000 [3]暨南大学,广东广州510000

出　　处：《中国兽医学报》2002年第5期430-432,共3页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目 ( 9710 0 10 9)

摘　　要：以 p CAT、p WB和 p WC为原始质粒 ,构建 WAP启动子调控的 Bcl- 2乳腺组织特异性表达载体 p WBS。载体用Bgl +Sal +Pvu 酶切 ,回收 3.0 kb的基因片段 WAP- Bcl- 2 - SV40 Poly A,通过显微注射方法导入 C5 7BL / 6 J×DBA/ 2 JF1代小鼠受精卵的雄原核内。共注射 10 0 0枚卵 ,将存活的约 6 0 0枚卵分别移植至 2 9只假孕母鼠输卵管内 ,获得仔鼠 5 5只。PCR检测 ,阳性鼠 12只。Southern blot检测 ,阳性鼠 4只 ,其中公、母鼠各 2只 ,在饲养过程中 ,2只公鼠均意外死亡。Western blot证实 ,1只母鼠 Bcl- 2表达阳性 ,其 F1代的 40只小鼠中 ,有 18只呈 PCR阳性 ,其中母鼠8只 ,在 5个月的反复怀孕过程中 ,未观察到乳房有任何肿瘤或肿块的出现 ,证实 Bcl- 2单独在转基因鼠乳腺中表达不会引起转基因鼠乳腺癌的发生。c-myc cDNA was cloned on the downstream of mouse WAP promoter to form a expression vector pWCS.Transgenic mice were established by microinjection of the 3.5 kb c-myc cDNA expression cassette purified from pWCS.600 ova were transplanted into twenty-nine pseudopregnant mouse.recipients.45 offspring were born from the ova mircroinjected.9 of them were proved to be gene integration positively by PCR,3 mice,1 male and 2 female,were further confirmed to be transgenic by Southern hybridization.The female mice died by accidently during its feeding.1 of the other 2 male mice could inherit the c-myc gene into its F1 generation proved by PCR.6 female of 35 mice in its F1 generation were comfirmed to be transgenic by PCR.They are at a stage of mutiple pregnancy and lactation.

关 键 词：人 Bcl-2乳腺 定位表达 转基因 小鼠 

分 类 号：Q78[生物学—分子生物学]